You searched for subject:(transfection).
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281 total matches.
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University of Alberta
1.
Wong, Peter Kwok Pan.
Exploring Novel Methods in Sonoportation.
Degree: MS, Department of Biomedical Engineering, 2012, University of Alberta
URL: https://era.library.ualberta.ca/files/2227mq96n 
► Sonoporation, a physical, non-viral, non-chemical transfection method, promises great potentials. However, many drawbacks hinders its generalization. Low transfection rate and cell viability after treatment are…
(more)
▼ Sonoporation, a physical, non-viral, non-chemical
transfection method, promises great potentials. However, many
drawbacks hinders its generalization. Low transfection rate and
cell viability after treatment are among the hindering factors of
sonoporation. The purpose of the research performed in this thesis
was to develop, explore and analyze new methodologies to overcome
the known drawbacks of sonoporation, which are to increase cell
viability and transfection rate. These novel methods include the
use of a self-developed ultrasound box, self-developed microbubble
carriers and the synergistic use of chemical transfection reagents.
Sonoporation were performed on MCF-7 and KG-1 cells as they
represent easy and difficult to transfect cell lines respectively.
Permeability markers, flow cytometry, MTT assay and MTS assay were
used to quantify transfection rate and cell viability after
sonoporation. New procedures were performed, analyzed and evaluated
for their feasibility for drug and or gene delivery. The thesis has
shown improvements in transfection rate and preserving viability.
However, sonoporation still remains an inefficient method to
deliver material into hard-to-transfect cells.
Subjects/Keywords: Delivery; Sonoporation; Transfection
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APA (6th Edition):
Wong, P. K. P. (2012). Exploring Novel Methods in Sonoportation. (Masters Thesis). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/2227mq96n
Chicago Manual of Style (16th Edition):
Wong, Peter Kwok Pan. “Exploring Novel Methods in Sonoportation.” 2012. Masters Thesis, University of Alberta. Accessed January 22, 2020.
https://era.library.ualberta.ca/files/2227mq96n.
MLA Handbook (7th Edition):
Wong, Peter Kwok Pan. “Exploring Novel Methods in Sonoportation.” 2012. Web. 22 Jan 2020.
Vancouver:
Wong PKP. Exploring Novel Methods in Sonoportation. [Internet] [Masters thesis]. University of Alberta; 2012. [cited 2020 Jan 22].
Available from: https://era.library.ualberta.ca/files/2227mq96n.
Council of Science Editors:
Wong PKP. Exploring Novel Methods in Sonoportation. [Masters Thesis]. University of Alberta; 2012. Available from: https://era.library.ualberta.ca/files/2227mq96n
Texas A&M University
2.
Mintzer, Meredith Ann.
The design and evaluation of triazine dendrimers for gene delivery.
Degree: 2011, Texas A&M University
URL: http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7330
► The interest in using gene therapy to target a variety of both inherited and acquired diseases has intensified over the last two decades. Because free…
(more)
▼ The interest in using gene therapy to target a variety of both inherited and
acquired diseases has intensified over the last two decades. Because free DNA is easily
degraded by serum nucleases in the bloodstream, the need for developing carrier systems
that can compact and protect the DNA was quickly realized. Viral vector systems were
some of the earliest carriers used, primarily because of the ease with which such systems
can infect host cells. However, difficulties experienced when using viral vectors,
including immunogenicity and the potential for genetic recombinations, forced
researchers to design alternative delivery strategies. Non-viral vectors offer one
alternative to overcome this dilemma. In addition to avoiding the biological problems
experienced using viral carriers, non-viral vectors also offer the potential for large-scale
production. Dendrimers are one non-viral carrier that has shown appreciable ability to
deliver DNA into cells, a process called
transfection. In the past, triazine dendrimers
have shown biocompatibility, and the ability to synthesize these structures to contain
cationic charges on the surface makes these structures potentially suitable for
transfection studies.
In this study, a small library of triazine dendrimers was synthesized in an attempt
to understand how variations to both the periphery and core of triazine dendrimers affect
the
transfection efficiency of these dendriplexes. In the first subset of structures, a
common core was used and various peripheral groups were appended to the dendrimer
surface. The physicochemical and biological data, obtained in collaboration with
Thomas Kissel at Philipps-Universitat Marburg, showed that the surface groups have a
notable affect on
transfection efficiency. Dendrimers with a higher amine number and
neutral surface groups show high DNA binding affinity and higher
transfection
efficiency. In the second subset of dendrimers, variations to the core showed that
transfection efficiency is improved both by increasing generation number and dendrimer
flexibility.
With this data in hand, triazine dendrimers with both higher generation number
and higher flexibility have been synthesized. Two different triazine linker groups,
trimethylene-dipiperidine and polyglycoldiamine, have been used. These structures will
be evaluated to determine if increasing both flexibility and generation number together
can further improve
transfection efficiency.
Advisors/Committee Members: Simanek, Eric E. (advisor), Bergbreiter, David E. (committee member), Singleton, Daniel A. (committee member), Grunlan, Melissa A. (committee member).
Subjects/Keywords: triazine; dendrimer; transfection; DNA
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APA (6th Edition):
Mintzer, M. A. (2011). The design and evaluation of triazine dendrimers for gene delivery. (Thesis). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7330
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Mintzer, Meredith Ann. “The design and evaluation of triazine dendrimers for gene delivery.” 2011. Thesis, Texas A&M University. Accessed January 22, 2020.
http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7330.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Mintzer, Meredith Ann. “The design and evaluation of triazine dendrimers for gene delivery.” 2011. Web. 22 Jan 2020.
Vancouver:
Mintzer MA. The design and evaluation of triazine dendrimers for gene delivery. [Internet] [Thesis]. Texas A&M University; 2011. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7330.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Mintzer MA. The design and evaluation of triazine dendrimers for gene delivery. [Thesis]. Texas A&M University; 2011. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2009-12-7330
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Texas A&M University
3.
Simeon, Rudo Lyndon.
Selecting a path against Hepatitis C Virus.
Degree: 2013, Texas A&M University
URL: http://hdl.handle.net/1969.1/151941
► Hepatitis C virus (HCV) currently affects ~5% of the world?s population and has relatively limited treatment options for infected patients. Genetic suppressor elements (GSE) derived…
(more)
▼ Hepatitis C virus (HCV) currently affects ~5% of the world?s population and has relatively limited treatment options for infected patients. Genetic suppressor elements (GSE) derived from a gene or genome of interest can act as transdominant inhibitors of a particular biology function presumably by binding to and blocking an essential interaction surface for protein activity. Taking advantage of hepatoma cell line n4mBid, that supports all stages of the HCV life cycle and strongly report HCV infection by a cell-death phenotype, we developed an iterative selection/enrichment strategy for the identification of GSE against HCV. Using this strategy, a library expressing random fragments of the HCV genome was screened for sequences able to suppress HCV infection. A 244 amino acid gene fragment, B1, was strongly enriched after 5 rounds of selection. B1 has a very high net positive charge of 43 at neutral pH and a high charge-to-mass (kDa) ratio of 1.5. We show that B1 expression specifically inhibits HCV replication, apparently due to its high positive charge. We also show that recombinant positively charged proteins can inhibit HCV infection, when supplied in vitro.In addition, eGFP-fused B1 potently penetrates both adherent and suspension cells with >80% of cells taking up the protein. Importantly, we show that B1 not only facilitates cellular uptake, but allows protein cargo to reach sites of biological relevance. B1 also delivers non-covalently conjugated RNA and DNA across the cell membrane to cytosolic and nuclear sites, with efficiency comparable to commercially available cationic lipid reagents. Our data suggest that B1 utilizes cell-surface glycans and multiple competing endocytic pathways to enter and traffic through cells.During a separate screening carried out in our lab, we identified a TACR3 inhibitor SB 222200 that had significant HCV activity. We go on to show that both TACR1 and TACR3 receptors are expressed in the HCV-permissive Huh 7.5 cell line. We also show that both TACR1 and TACR3 inhibitors significantly inhibit HCV infection. These results point to the potential for TACR1 antagonists in treating patients infected with both HCV and HIV.
Advisors/Committee Members: Chen, Zhilei (advisor), Jayaraman, Arul (committee member), Karim, Nazmul (committee member), Leibowitz, Julian (committee member).
Subjects/Keywords: HCV; biomolecules; biotherapeutics; transfection; transduction
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APA (6th Edition):
Simeon, R. L. (2013). Selecting a path against Hepatitis C Virus. (Thesis). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/151941
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Simeon, Rudo Lyndon. “Selecting a path against Hepatitis C Virus.” 2013. Thesis, Texas A&M University. Accessed January 22, 2020.
http://hdl.handle.net/1969.1/151941.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Simeon, Rudo Lyndon. “Selecting a path against Hepatitis C Virus.” 2013. Web. 22 Jan 2020.
Vancouver:
Simeon RL. Selecting a path against Hepatitis C Virus. [Internet] [Thesis]. Texas A&M University; 2013. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/1969.1/151941.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Simeon RL. Selecting a path against Hepatitis C Virus. [Thesis]. Texas A&M University; 2013. Available from: http://hdl.handle.net/1969.1/151941
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Hong Kong
4.
蕭景新.; Siu, King-sun.
Development of crosslinked poly(ethylenimine) as a
potential nucleic acid delivery agent.
Degree: M. Phil., 2009, University of Hong Kong
URL: Siu,
K.
[蕭景新].
(2009).
Development
of
crosslinked
poly(ethylenimine)
as
a
potential
nucleic
acid
delivery
agent.
(Thesis).
University
of
Hong
Kong,
Pokfulam,
Hong
Kong
SAR.
Retrieved
from
http://dx.doi.org/10.5353/th_b4322387
;
http://dx.doi.org/10.5353/th_b4322387
;
http://hdl.handle.net/10722/130725
published_or_final_version
Chemistry
Master
Master of Philosophy
Advisors/Committee Members: Lin, MC.Subjects/Keywords: Polymers.; Transfection.
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APA ·
Chicago ·
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CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
蕭景新.; Siu, K. (2009). Development of crosslinked poly(ethylenimine) as a
potential nucleic acid delivery agent. (Masters Thesis). University of Hong Kong. Retrieved from Siu, K. [蕭景新]. (2009). Development of crosslinked poly(ethylenimine) as a potential nucleic acid delivery agent. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4322387 ; http://dx.doi.org/10.5353/th_b4322387 ; http://hdl.handle.net/10722/130725
Chicago Manual of Style (16th Edition):
蕭景新.; Siu, King-sun. “Development of crosslinked poly(ethylenimine) as a
potential nucleic acid delivery agent.” 2009. Masters Thesis, University of Hong Kong. Accessed January 22, 2020.
Siu, K. [蕭景新]. (2009). Development of crosslinked poly(ethylenimine) as a potential nucleic acid delivery agent. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4322387 ; http://dx.doi.org/10.5353/th_b4322387 ; http://hdl.handle.net/10722/130725.
MLA Handbook (7th Edition):
蕭景新.; Siu, King-sun. “Development of crosslinked poly(ethylenimine) as a
potential nucleic acid delivery agent.” 2009. Web. 22 Jan 2020.
Vancouver:
蕭景新.; Siu K. Development of crosslinked poly(ethylenimine) as a
potential nucleic acid delivery agent. [Internet] [Masters thesis]. University of Hong Kong; 2009. [cited 2020 Jan 22].
Available from: Siu, K. [蕭景新]. (2009). Development of crosslinked poly(ethylenimine) as a potential nucleic acid delivery agent. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4322387 ; http://dx.doi.org/10.5353/th_b4322387 ; http://hdl.handle.net/10722/130725.
Council of Science Editors:
蕭景新.; Siu K. Development of crosslinked poly(ethylenimine) as a
potential nucleic acid delivery agent. [Masters Thesis]. University of Hong Kong; 2009. Available from: Siu, K. [蕭景新]. (2009). Development of crosslinked poly(ethylenimine) as a potential nucleic acid delivery agent. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4322387 ; http://dx.doi.org/10.5353/th_b4322387 ; http://hdl.handle.net/10722/130725
University of Hong Kong
5.
Lai, Wing-fu.
Fabrication and properties of poly(ethylenimine)-based
polymers for non-viral plasmid delivery.
Degree: PhD, 2011, University of Hong Kong
URL: Lai,
W.
[黎永富].
(2011).
Fabrication
and
properties
of
poly(ethylenimine)-based
polymers
for
non-viral
plasmid
delivery.
(Thesis).
University
of
Hong
Kong,
Pokfulam,
Hong
Kong
SAR.
Retrieved
from
http://dx.doi.org/10.5353/th_b4658812
;
http://dx.doi.org/10.5353/th_b4658812
;
http://hdl.handle.net/10722/143215
published_or_final_version
Chemistry
Doctoral
Doctor of Philosophy
Advisors/Committee Members: Sun, H.Subjects/Keywords: Transfection.; Polymers.
Record Details
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APA ·
Chicago ·
MLA ·
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CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Lai, W. (2011). Fabrication and properties of poly(ethylenimine)-based
polymers for non-viral plasmid delivery. (Doctoral Dissertation). University of Hong Kong. Retrieved from Lai, W. [黎永富]. (2011). Fabrication and properties of poly(ethylenimine)-based polymers for non-viral plasmid delivery. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4658812 ; http://dx.doi.org/10.5353/th_b4658812 ; http://hdl.handle.net/10722/143215
Chicago Manual of Style (16th Edition):
Lai, Wing-fu. “Fabrication and properties of poly(ethylenimine)-based
polymers for non-viral plasmid delivery.” 2011. Doctoral Dissertation, University of Hong Kong. Accessed January 22, 2020.
Lai, W. [黎永富]. (2011). Fabrication and properties of poly(ethylenimine)-based polymers for non-viral plasmid delivery. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4658812 ; http://dx.doi.org/10.5353/th_b4658812 ; http://hdl.handle.net/10722/143215.
MLA Handbook (7th Edition):
Lai, Wing-fu. “Fabrication and properties of poly(ethylenimine)-based
polymers for non-viral plasmid delivery.” 2011. Web. 22 Jan 2020.
Vancouver:
Lai W. Fabrication and properties of poly(ethylenimine)-based
polymers for non-viral plasmid delivery. [Internet] [Doctoral dissertation]. University of Hong Kong; 2011. [cited 2020 Jan 22].
Available from: Lai, W. [黎永富]. (2011). Fabrication and properties of poly(ethylenimine)-based polymers for non-viral plasmid delivery. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4658812 ; http://dx.doi.org/10.5353/th_b4658812 ; http://hdl.handle.net/10722/143215.
Council of Science Editors:
Lai W. Fabrication and properties of poly(ethylenimine)-based
polymers for non-viral plasmid delivery. [Doctoral Dissertation]. University of Hong Kong; 2011. Available from: Lai, W. [黎永富]. (2011). Fabrication and properties of poly(ethylenimine)-based polymers for non-viral plasmid delivery. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4658812 ; http://dx.doi.org/10.5353/th_b4658812 ; http://hdl.handle.net/10722/143215
Jawaharlal Nehru University
6.
Prasad, T. K.
Cell biology of lipid mediated transfection;.
Degree: Centre for Cellular and Molecular
Biology, 2004, Jawaharlal Nehru University
URL: http://shodhganga.inflibnet.ac.in/handle/10603/16453
► Gene delivery is a process by which an exogenous copy of the gene or DNA is newlineintroduced into the cell. Gene delivery has tremendous medical…
(more)
▼ Gene delivery is a process by which an exogenous
copy of the gene or DNA is newlineintroduced into the cell. Gene
delivery has tremendous medical implications in newlinetreating
several genetic disorders, which are refractory to the conventional
newlinemedicines. Several strategies have been developed to
introduce the foreign DNA newlineinto the cells. Among the most
efficient ones are viruses, which due to their newlinenatural
ability to carry the genetic material into the cells have attracted
scientists newlineto use them to carry DNA fragment of our interest
into the cells. However, due to newlinetheir harmful potential as
well as safety concerns, other alternative non-viral newlinemethods
have been developed and tested in the recent years to deliver DNA
newlinemuch more safely. One promising vehicle among such non-viral
categories is newlinecationic liposomes. Cationic lipids have
gained popularity because of the several newlineadvantages that
they have shown over viral based methods.
Bibliography p.90-105
Advisors/Committee Members: Rao, N. Madhusudhana.
Subjects/Keywords: Cell biology; mediated transfection
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APA (6th Edition):
Prasad, T. K. (2004). Cell biology of lipid mediated transfection;. (Thesis). Jawaharlal Nehru University. Retrieved from http://shodhganga.inflibnet.ac.in/handle/10603/16453
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Prasad, T K. “Cell biology of lipid mediated transfection;.” 2004. Thesis, Jawaharlal Nehru University. Accessed January 22, 2020.
http://shodhganga.inflibnet.ac.in/handle/10603/16453.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Prasad, T K. “Cell biology of lipid mediated transfection;.” 2004. Web. 22 Jan 2020.
Vancouver:
Prasad TK. Cell biology of lipid mediated transfection;. [Internet] [Thesis]. Jawaharlal Nehru University; 2004. [cited 2020 Jan 22].
Available from: http://shodhganga.inflibnet.ac.in/handle/10603/16453.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Prasad TK. Cell biology of lipid mediated transfection;. [Thesis]. Jawaharlal Nehru University; 2004. Available from: http://shodhganga.inflibnet.ac.in/handle/10603/16453
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Rochester Institute of Technology
7.
Ahmed, Adeel.
Efficient Template-Based Nanomanufacturing of Carbon Nanotube Arrays for Cell Applications.
Degree: MS, Mechanical Engineering, 2017, Rochester Institute of Technology
URL: https://scholarworks.rit.edu/theses/9524
► Carbon nanotube arrays have been found to be highly effective at carrying out intracellular delivery of cargo at high efficiencies while ensuring cell viability.…
(more)
▼ Carbon nanotube arrays have been found to be highly effective at carrying out intracellular delivery of cargo at high efficiencies while ensuring cell viability. Template based chemical vapor deposition is a commonly used process to fabricate these arrays. However, current etching methods used to expose carbon nanotubes from templates are expensive and time consuming. The high cost and time-consuming processes currently required to fabricate such arrays are factors which limit the commercialization of this technology and inhibit scope for larger research programs. In this thesis, alternative nanofabrication methods were explored with the aim of making the fabrication of CNT arrays cost effective and efficient. Mechanical polishing coupled with wet chemical etching is shown as a feasible alternative option to dry etching. The effects of process variables on physical properties of CNT arrays have been studied and quantified in order to demonstrate control over the process. Scanning Electron Microscopy has been used to qualitatively understand the differences between CNT arrays fabricated using dry etching and the alternative process. Cell culture has been demonstrated on the CNT arrays and the potential to use 3D printing to fabricate a nanofluidic device is also demonstrated. The alternative process can save etching time by 97% while maintaining a similar level of control over the process. This study, therefore, opens the path to quicker production of CNT arrays at low cost for biomedical use.
Advisors/Committee Members: Michael G. Schrlau, Kathleen Lamkin-Kennard, Thomas Gaborski.
Subjects/Keywords: Carbon; Membrane; Nanofabrication; Nanotube; Transfection
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APA ·
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APA (6th Edition):
Ahmed, A. (2017). Efficient Template-Based Nanomanufacturing of Carbon Nanotube Arrays for Cell Applications. (Masters Thesis). Rochester Institute of Technology. Retrieved from https://scholarworks.rit.edu/theses/9524
Chicago Manual of Style (16th Edition):
Ahmed, Adeel. “Efficient Template-Based Nanomanufacturing of Carbon Nanotube Arrays for Cell Applications.” 2017. Masters Thesis, Rochester Institute of Technology. Accessed January 22, 2020.
https://scholarworks.rit.edu/theses/9524.
MLA Handbook (7th Edition):
Ahmed, Adeel. “Efficient Template-Based Nanomanufacturing of Carbon Nanotube Arrays for Cell Applications.” 2017. Web. 22 Jan 2020.
Vancouver:
Ahmed A. Efficient Template-Based Nanomanufacturing of Carbon Nanotube Arrays for Cell Applications. [Internet] [Masters thesis]. Rochester Institute of Technology; 2017. [cited 2020 Jan 22].
Available from: https://scholarworks.rit.edu/theses/9524.
Council of Science Editors:
Ahmed A. Efficient Template-Based Nanomanufacturing of Carbon Nanotube Arrays for Cell Applications. [Masters Thesis]. Rochester Institute of Technology; 2017. Available from: https://scholarworks.rit.edu/theses/9524
University of British Columbia
8.
Hogge, Donna Eileen.
Genetic investigations of human hemopoiesis : studies of clonality and gene transfer to hemopoietic progenitors
.
Degree: 1987, University of British Columbia
URL: http://hdl.handle.net/2429/27316
► In most neoplasms malignant change occurs in a single cell which then proliferates. My purpose was to explore methods to study the cell that gives…
(more)
▼ In most neoplasms malignant change occurs in a single cell which then proliferates. My purpose was to explore methods to study the cell that gives rise to hemopoietic cancer and to investigate the abnormalities at a molecular level.
Cytogenetic analysis of cells from individual hemopoietic colonies revealed that monosomy 7 syndrome, a hematologic disorder of childhood, arises in a primitive cell capable of differentiating down both myeloid and erythroid pathways.
Long-term bone marrow cultures (LTC) from patients with chronic myelogenous leukemia (CML) favor the growth of Philadelphia chromosome (Ph) negative progenitors which, although cytogenetically normal, could have been part of the malignant clone at a stage prior to the development of the Ph. LTC's were initiated with cells from 2 women with CML who were heterozygous for 2 electrophoretically distinct glucose-6-phosphate dehydrogenase (G6PD) enzyme variants. In one patient, 2/11 progenitors were Ph-negative after 4 to 6 weeks in LTC and 4/30 were nonclonal by G6PD enzyme analysis, i.e. the colonies expressed the enzyme not found in the malignant clone. In this case, karyotypically normal cells were truly normal.
Next, gene transfer to human hemopoietic cells was demonstrated using recombinant retrovirus carrying the selectable marker gene, neor. With the K562 human leukemic cell line as targets up to 60% of infected cells became G418 resistant (G418r). Cloned populations of G418r cells showed unique patterns of retroviral integration in K562 DNA. When the target cells were progenitors from normal marrow, CML blood or fetal liver, the highest frequencies of G418r granulocyte-macrophage or large erythroid colonies was 16% and 5% respectively. Experiments infecting bone marrow cells in LTC with neor virus produced up to 2% G418r colonies after as long as 3 weeks in culture. Using v-src virus to infect LTC failed to perturb hemopoiesis, although infection of bone marrow-derived cells in these cultures was documented.
In summary:
1. Unique populations of hemopoietic progenitors can be identified in culture using several genetic markers including chromosomes, G6PD analysis or gene transfer. 2. The feasibility of retroviral-mediated gene transfer for use on human hemopoietic cells has been demonstrated.
Subjects/Keywords: Transfection;
Hematopoiesis
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APA ·
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Manager
APA (6th Edition):
Hogge, D. E. (1987). Genetic investigations of human hemopoiesis : studies of clonality and gene transfer to hemopoietic progenitors
. (Thesis). University of British Columbia. Retrieved from http://hdl.handle.net/2429/27316
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Hogge, Donna Eileen. “Genetic investigations of human hemopoiesis : studies of clonality and gene transfer to hemopoietic progenitors
.” 1987. Thesis, University of British Columbia. Accessed January 22, 2020.
http://hdl.handle.net/2429/27316.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Hogge, Donna Eileen. “Genetic investigations of human hemopoiesis : studies of clonality and gene transfer to hemopoietic progenitors
.” 1987. Web. 22 Jan 2020.
Vancouver:
Hogge DE. Genetic investigations of human hemopoiesis : studies of clonality and gene transfer to hemopoietic progenitors
. [Internet] [Thesis]. University of British Columbia; 1987. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/2429/27316.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Hogge DE. Genetic investigations of human hemopoiesis : studies of clonality and gene transfer to hemopoietic progenitors
. [Thesis]. University of British Columbia; 1987. Available from: http://hdl.handle.net/2429/27316
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Texas State University – San Marcos
9.
Palacios, Adriana S.
Transfection Efficiencies of Chitosan/Sirna Nanoparticles in Colorectal Cancer Cells.
Degree: MS, Biochemistry, 2012, Texas State University – San Marcos
URL: https://digital.library.txstate.edu/handle/10877/5487
► RNA interference (RNAi) is a recently discovered phenomenon that employs the use of miRNA (microRNA) and siRNA (short interfering RNA) that can be utilized for…
(more)
▼ RNA interference (RNAi) is a recently discovered phenomenon that employs the use of miRNA (microRNA) and siRNA (short interfering RNA) that can be utilized for gene therapy purposes. However, the problem that has arisen is delivery of the siRNAs into diseased cells. The compound chitosan has recently gained much attention because it possesses several properties that make it an ideal candidate as a delivery vehicle for siRNA. Chitosan/siRNA based nanoparticles have been synthesized and characterized that are less than 100 nm in diameter. Characterization of their size was completed by using transmission electron microscopy (TEM), scanning electron microscopy (SEM), and dynamic light scattering (DLS) techniques. Characterization of siRNA loading was accomplished by utilizing UV spectroscopy, EDS, and confocal microscopy to test encapsulation of siRNA. The colorectal cancer cell line HCT 116, was used to test
transfection of siRNA across the cellular membrane. The
transfection efficiencies were evaluated by using fluorescently tagged siRNA embedded within chitosan nanoparticles. The chitosan/siRNA delivery system was compared with the classical approach used for siRNA delivery; namely, siRNA in Lipofectamine 2000 as well as with cells that received no treatment. Live cell imaging was also employed to aid in visualization of
transfection. The results show that the chitosan based systems do transfect siRNA across the cellular member.
Advisors/Committee Members: Rudzinski, Walter E. (advisor), Whitten, Steven T. (committee member), Betancourt, Tania (committee member).
Subjects/Keywords: Transfection efficiency; siRNA; Chitosan nanoparticles
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APA (6th Edition):
Palacios, A. S. (2012). Transfection Efficiencies of Chitosan/Sirna Nanoparticles in Colorectal Cancer Cells. (Masters Thesis). Texas State University – San Marcos. Retrieved from https://digital.library.txstate.edu/handle/10877/5487
Chicago Manual of Style (16th Edition):
Palacios, Adriana S. “Transfection Efficiencies of Chitosan/Sirna Nanoparticles in Colorectal Cancer Cells.” 2012. Masters Thesis, Texas State University – San Marcos. Accessed January 22, 2020.
https://digital.library.txstate.edu/handle/10877/5487.
MLA Handbook (7th Edition):
Palacios, Adriana S. “Transfection Efficiencies of Chitosan/Sirna Nanoparticles in Colorectal Cancer Cells.” 2012. Web. 22 Jan 2020.
Vancouver:
Palacios AS. Transfection Efficiencies of Chitosan/Sirna Nanoparticles in Colorectal Cancer Cells. [Internet] [Masters thesis]. Texas State University – San Marcos; 2012. [cited 2020 Jan 22].
Available from: https://digital.library.txstate.edu/handle/10877/5487.
Council of Science Editors:
Palacios AS. Transfection Efficiencies of Chitosan/Sirna Nanoparticles in Colorectal Cancer Cells. [Masters Thesis]. Texas State University – San Marcos; 2012. Available from: https://digital.library.txstate.edu/handle/10877/5487
University of Minnesota
10.
Boyle, William.
Charged Glycopolymer Materials for Epidermolysis Bullosa.
Degree: PhD, Chemistry, 2017, University of Minnesota
URL: http://hdl.handle.net/11299/201073
► Improved delivery of therapeutic nucleic acid payloads to cells could lead to dramatically improved clinical outcomes for patients suffering from genetic disorders. This work focuses…
(more)
▼ Improved delivery of therapeutic nucleic acid payloads to cells could lead to dramatically improved clinical outcomes for patients suffering from genetic disorders. This work focuses on the use of a trehalose-containing cationic glycopolymer, termed Tr4, to transfect clinically relevant cell types. In particular, the development of gene delivery methods to improve the transfection of cell types associated with the skin disease epidermolysis bullosa are investigated. The sulfated glycosaminoglycan, heparin, is show to form ternary complexes with pDNA and Tr4 leading to dramatically increased transfection efficiency in primary fibroblasts, induced pluripotent stem cells, HepG2, and U87-MG cells. This increase is not caused by improved uptake, but instead appears to be driven by improved intracellular trafficking of polyplexes compared transfection without heparin. Increasing the size of the plasmid cargo from 4.7 kbp to a more therapeutically relevant 10 kbp leads to the complete loss of transfection efficiency in Tr4-heparin transfection of primary fibroblasts and a reduction in transfection efficiency in iPSCs. Co-transfecting with additives meant to increase nuclear localization of the pDNA recovers the efficiency lost by increasing the plasmid size. These techniques allowed for the development of function transfection methods in iPSCs delivering a synthetic transcription activator of collagen type VII. Finally, nanofiber mats containing chondroitin sulfate were developed to scavenge inflammatory molecules from wound exudate.
Subjects/Keywords: Epidermolysis Bullosa; Gene Delivery; Polymers; Transfection
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APA (6th Edition):
Boyle, W. (2017). Charged Glycopolymer Materials for Epidermolysis Bullosa. (Doctoral Dissertation). University of Minnesota. Retrieved from http://hdl.handle.net/11299/201073
Chicago Manual of Style (16th Edition):
Boyle, William. “Charged Glycopolymer Materials for Epidermolysis Bullosa.” 2017. Doctoral Dissertation, University of Minnesota. Accessed January 22, 2020.
http://hdl.handle.net/11299/201073.
MLA Handbook (7th Edition):
Boyle, William. “Charged Glycopolymer Materials for Epidermolysis Bullosa.” 2017. Web. 22 Jan 2020.
Vancouver:
Boyle W. Charged Glycopolymer Materials for Epidermolysis Bullosa. [Internet] [Doctoral dissertation]. University of Minnesota; 2017. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/11299/201073.
Council of Science Editors:
Boyle W. Charged Glycopolymer Materials for Epidermolysis Bullosa. [Doctoral Dissertation]. University of Minnesota; 2017. Available from: http://hdl.handle.net/11299/201073
University of Houston
11.
Hoang, Johnson 1988-.
Mixed Quaternary-Ammonium-Coated Gold Nanoparticles for Delivery of MicroRNA.
Degree: Biology and Biochemistry, Department of, 2018, University of Houston
URL: http://hdl.handle.net/10657/3425
► The research described in this dissertation seeks to develop an inorganic transfection delivery carrier using a custom-designed self-assembled monolayer (SAM). Initial studies describe the preparation…
(more)
▼ The research described in this dissertation seeks to develop an inorganic
transfection delivery carrier using a custom-designed self-assembled monolayer (SAM). Initial studies describe the preparation of SAMs on flat gold surfaces derived from three unique bidentate adsorbates and two analogous monodentate adsorbates of different chain lengths bearing a quaternary-ammonium terminal group. The generated SAMs were evaluated using ellipsometry, X-ray photoelectron spectroscopy (XPS), contact angle goniometry, polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS), and electrochemical quartz crystal microbalance (eQCM). The effect of the packing density of the SAMs on oligonucleotide binding was evaluated in regards to the terminal group used. In an effort to enhance the immobilization of oligonucleotides on the SAMs surfaces, the use of a mixed monolayer containing a long and short-chained adsorbate was utilized. The SAM surface comprised of a 50:50 ratio was determined to have the greatest amount of immobilized oligonucleotides.
To further expand on the initial study and to design a nanocarrier capable of initiating the "proton sponge" effect in cells, the quaternary-ammonium-terminated adsorbate was mixed with an ammonium-terminated adsorbate, both bearing a bidentate headgroup. The effect of burying either the quaternary-ammonium or ammonium moiety into the film on oligonucleotide loading was investigated. Based on our analysis, the SAM surface comprised of a 75:25 (ammonium: quaternary-ammonium) in which the quaternary ammonium species is buried resulted in the highest oligonucleotide loading.
The development of a layer-by-layer gold nanoparticle complex for the delivery of miRNA was pursued using the 75:25 SAM surface. The nanocomplex provided a platform to electrostatically bind miRNA through its phosphate backbone. The uniquely designed layer-by-layer gold nanocomplex was characterized by dynamic light scattering (DLS), gel retardation assay, transmission electron microscope (TEM), ultraviolet visible spectroscopy (UV-vis), X-ray photoelectron spectroscopy (XPS), and zeta potential (ζ-potential). Furthermore, miRNA immobilized onto the gold nanocomplex was transfected into osteosarcoma (HOS) cell lines and then quantified by quantitative polymerase chain reaction (qPCR). The cytotoxicity of the nanocomplex was then examined by MTS assay. Overall, the gold nanocomplex was successfully developed and shown to have low toxicity while delivering miRNA into an HOS cell line with a 10,000-fold increase.
Advisors/Committee Members: Gunaratne, Preethi H (advisor), Lee, T. Randall (committee member), Bark, Steven (committee member), Bartholomeusz, Geoffrey A (committee member), Fox, Robert (committee member).
Subjects/Keywords: Gold Nanoparticles; Self-Assembled Monolayer; Nanocarrier; Transfection
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APA (6th Edition):
Hoang, J. 1. (2018). Mixed Quaternary-Ammonium-Coated Gold Nanoparticles for Delivery of MicroRNA. (Thesis). University of Houston. Retrieved from http://hdl.handle.net/10657/3425
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Hoang, Johnson 1988-. “Mixed Quaternary-Ammonium-Coated Gold Nanoparticles for Delivery of MicroRNA.” 2018. Thesis, University of Houston. Accessed January 22, 2020.
http://hdl.handle.net/10657/3425.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Hoang, Johnson 1988-. “Mixed Quaternary-Ammonium-Coated Gold Nanoparticles for Delivery of MicroRNA.” 2018. Web. 22 Jan 2020.
Vancouver:
Hoang J1. Mixed Quaternary-Ammonium-Coated Gold Nanoparticles for Delivery of MicroRNA. [Internet] [Thesis]. University of Houston; 2018. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/10657/3425.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Hoang J1. Mixed Quaternary-Ammonium-Coated Gold Nanoparticles for Delivery of MicroRNA. [Thesis]. University of Houston; 2018. Available from: http://hdl.handle.net/10657/3425
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Boston University
12.
Hayward, Oliver James.
Mechanisms of Mononegavirales gene expression.
Degree: MS, Medical Sciences, 2019, Boston University
URL: http://hdl.handle.net/2144/38661
► The Mononegavirales order unifies the non-segmented negative sense viruses (nsNSVs), including Marburgvirus (MARV) of the Filoviridae family and respiratory syncytial virus (RSV) of the Pneumoviridae.…
(more)
▼ The Mononegavirales order unifies the non-segmented negative sense viruses (nsNSVs), including Marburgvirus (MARV) of the Filoviridae family and respiratory syncytial virus (RSV) of the Pneumoviridae. The mechanism of action of these viruses and how they infect cells are very similar, especially when focusing on their polymerases, which are distinct from those of eukaryotes and therefore possible targets for antiviral drugs. nsNSVs utilize a RNA-dependent RNA polymerase to either replicate the viral RNA genome or transcribe it into positive sense mRNA. Despite this, these two viruses result in very different, but equally devastating, effects in humans. Whereas MARV virus often results in rare but fatal hemorrhagic fevers, RSV is a common seasonal virus that can result in long term negative effects to respiratory health. These negative effects on public health demand extensive research in these two fields and a need to develop new technology and methods in order to uncover the missing pieces of viral gene expression. Specifically, the development of a MARV minigenome system would allow for increased testing of this virus outside of the confines of the biosafety level 4 (BSL-4) setting. By replacing MARV genes with reporter genes, but retaining the characteristic leader, intergenic, and trailer regions of the genome, tests involving site directed mutagenesis would reveal new insights into the crucial genomic elements needed for successful gene expression. Coupled with the
transfection of the minigenome with plasmids coding for the crucial MARV proteins, artificial changes to the genome would lead to the presence of absence of translated bioluminescent reporter proteins. Using these two viruses, this study attempted to find commonalities across families. Specifically, the goals of this research were twofold, to find the optimal ratio of MARV plasmids in the minigenome system to understand the effects of the stem-loop secondary structure of MARV mRNA transcripts as well as determine the tail length of the poly(A) tail of RSV mRNA transcripts using digestion and probing primers. Calculating the RSV poly(A) tail length would allow for further research into determining whether the MARV and RSV polymerase polyadenylates before or after it releases the transcript. Despite multiple failed attempts, transfections using pCAGGS plasmids and the eGFP monocistronic minigenome in a 6-well plate qualitatively demonstrated the need for pCAGGS-L plasmid concentration of 1000 ng/µl. Due to time constraints, the poly(A) tail length of the RSV NS-1 mRNA transcript could not be determined. Overall, this study focused on gaining new insights on the techniques and procedures necessary for conducting virus research in a biosafety level 2 (BSL-2) setting, as well as developing troubleshooting skills in approaching fail experiments.
Advisors/Committee Members: Fearns, Rachel (advisor), Muhlberger, Elke (advisor).
Subjects/Keywords: Medicine; Marburg; Minigenome; Polyadenylation; Respiratory; Syncytial; Transfection
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APA (6th Edition):
Hayward, O. J. (2019). Mechanisms of Mononegavirales gene expression. (Masters Thesis). Boston University. Retrieved from http://hdl.handle.net/2144/38661
Chicago Manual of Style (16th Edition):
Hayward, Oliver James. “Mechanisms of Mononegavirales gene expression.” 2019. Masters Thesis, Boston University. Accessed January 22, 2020.
http://hdl.handle.net/2144/38661.
MLA Handbook (7th Edition):
Hayward, Oliver James. “Mechanisms of Mononegavirales gene expression.” 2019. Web. 22 Jan 2020.
Vancouver:
Hayward OJ. Mechanisms of Mononegavirales gene expression. [Internet] [Masters thesis]. Boston University; 2019. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/2144/38661.
Council of Science Editors:
Hayward OJ. Mechanisms of Mononegavirales gene expression. [Masters Thesis]. Boston University; 2019. Available from: http://hdl.handle.net/2144/38661
University of Ottawa
13.
Cencen, Veronika.
A Microfluidic Device for Transfection of Mammalian Cells Using Adjustable Shear Stress
.
Degree: 2016, University of Ottawa
URL: http://hdl.handle.net/10393/35525
► Microfluidic technology is a rapidly progressing tool in biomedical engineering. Microfluidic devices are appreciated for their simplicity and production efficiency potential. Our research focuses on…
(more)
▼ Microfluidic technology is a rapidly progressing tool in biomedical engineering. Microfluidic devices are appreciated for their simplicity and production efficiency potential. Our research focuses on developing a microfluidic device capable of transfecting cells by applying shear stress to cause temporary membrane damage. The advantage of this physical method of transfection is the possibility of incorporating large molecules that cannot be inserted with more traditional chemical transfection methods, while avoiding the large fall in viability seen with other physical methods such as electroporation. Unlike previous groups, our device incorporates the use of microfluidic valves to allow tunability, and impedance sensing for cell membrane damage analysis. We achieve (95±5)% cell viability and up to (68±5)% efficiency in transfecting 3T3 cells with DNA-sized molecules. In future stages, we intend to add the device onto an existing cell-encapsulation device that is tasked with preparing therapeutic cells to be used in regenerative medicine applications.
Subjects/Keywords: Microfluidics;
Transfection;
Impedance;
Photolithograpy;
PAH;
FITC
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APA (6th Edition):
Cencen, V. (2016). A Microfluidic Device for Transfection of Mammalian Cells Using Adjustable Shear Stress
. (Thesis). University of Ottawa. Retrieved from http://hdl.handle.net/10393/35525
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Cencen, Veronika. “A Microfluidic Device for Transfection of Mammalian Cells Using Adjustable Shear Stress
.” 2016. Thesis, University of Ottawa. Accessed January 22, 2020.
http://hdl.handle.net/10393/35525.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Cencen, Veronika. “A Microfluidic Device for Transfection of Mammalian Cells Using Adjustable Shear Stress
.” 2016. Web. 22 Jan 2020.
Vancouver:
Cencen V. A Microfluidic Device for Transfection of Mammalian Cells Using Adjustable Shear Stress
. [Internet] [Thesis]. University of Ottawa; 2016. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/10393/35525.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Cencen V. A Microfluidic Device for Transfection of Mammalian Cells Using Adjustable Shear Stress
. [Thesis]. University of Ottawa; 2016. Available from: http://hdl.handle.net/10393/35525
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Minnesota
14.
Phillips, Haley.
Elucidating Glycopolycation Structure-Function Relationships For Improved Gene Therapy.
Degree: PhD, Chemistry, 2017, University of Minnesota
URL: http://hdl.handle.net/11299/206410
► The gene therapy field is devoted to treating disease by adding, altering, or inhibiting gene expression. This type of therapy holds great promise for the…
(more)
▼ The gene therapy field is devoted to treating disease by adding, altering, or inhibiting gene expression. This type of therapy holds great promise for the treatment and even cure of monogenic diseases such as cystic fibrosis, Duchenne muscular dystrophy, hemophilia A and B, and epidermolysis bullosa. To produce therapeutic effect, nucleic acids must be delivered and expressed in cells of interest. Deoxyribonucleic acids (DNA) and ribonucleic acids (RNA) in many forms can be delivered using viral or non-viral vehicles. Viral vectors provide efficient DNA delivery; however, packaging limitations and occasional safety issues such as immune responses are major issues. In contrast, non-viral vectors are cheaper and easier to mass produce and can package any length of nucleic acid; however, non-viral vectors struggle to deliver genetic cargo at therapeutically beneficial levels. Polymers with the ability to condense and protect genetic material make promising non-viral vectors. They are relatively easy to produce compared to viral vehicles, can safely package various plasmid sizes, and have shown significant uptake in a wide variety of human cell lines. Cationic polymers complex with the negatively charged phosphodiester backbone of DNA or RNA, forming inter-polyelectrolyte complexes termed polyplexes. Herein, we explore using experiments in vitro, ex vivo, and in vivo to probe the structure-function relationships dictating polyplex gene delivery and other glycomaterial applications.
Subjects/Keywords: Biocompatibility; Delivery; Glycopolymer; Nonviral; Polyplex; Transfection
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APA (6th Edition):
Phillips, H. (2017). Elucidating Glycopolycation Structure-Function Relationships For Improved Gene Therapy. (Doctoral Dissertation). University of Minnesota. Retrieved from http://hdl.handle.net/11299/206410
Chicago Manual of Style (16th Edition):
Phillips, Haley. “Elucidating Glycopolycation Structure-Function Relationships For Improved Gene Therapy.” 2017. Doctoral Dissertation, University of Minnesota. Accessed January 22, 2020.
http://hdl.handle.net/11299/206410.
MLA Handbook (7th Edition):
Phillips, Haley. “Elucidating Glycopolycation Structure-Function Relationships For Improved Gene Therapy.” 2017. Web. 22 Jan 2020.
Vancouver:
Phillips H. Elucidating Glycopolycation Structure-Function Relationships For Improved Gene Therapy. [Internet] [Doctoral dissertation]. University of Minnesota; 2017. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/11299/206410.
Council of Science Editors:
Phillips H. Elucidating Glycopolycation Structure-Function Relationships For Improved Gene Therapy. [Doctoral Dissertation]. University of Minnesota; 2017. Available from: http://hdl.handle.net/11299/206410
Duke University
15.
Wu, Mina.
Implicated Role of Endocytosis in the Internalization and Intracellular Transport of Plasmid DNA During Electric Field-Mediated Gene Delivery
.
Degree: 2011, Duke University
URL: http://hdl.handle.net/10161/4994
► Electric field mediated gene delivery (EFMGD) or electrotransfection is a popular, non-viral gene delivery method that has been used in a variety of studies…
(more)
▼ Electric field mediated gene delivery (EFMGD) or electrotransfection is a popular, non-viral gene delivery method that has been used in a variety of studies and applications ranging from basic cell biology research to clinical gene therapy. Yet, the mechanism(s) by which electrotransfection facilitates DNA delivery across the cell membrane into the cell and its subsequent intracellular transport across the cytosolic space towards the nucleus have been insufficiently studied and still remain controversial. Understanding these mechanisms and characterizing the intracellular journey of pDNA is important for understanding the physiological barriers of EFMGD within the cell, which can be used to engineer better solutions to overcome these barriers with the ultimate goal of improving the
transfection efficiency of this technology. Conventional thought in the field assumes that such transport modes as diffusion, electrophoresis, and electro-osmosis, which govern the entry of small molecules into cells through electric field-generated transient membrane pores, also apply to electric field-mediated delivery of therapeutic DNA. We propose that electrically-induced gene transfer into cells is governed by an alternative, more active mode of transport that entails the involvement of cellular endocytic processes. It is our hypothesis that pulsed electric field generate these membrane pores which interact with nearby DNA molecules; but that actual DNA translocation across the membrane is driven by endocytosis, which consequently, then, also plays a role in the intracellular transport of the DNA. To this end, we first investigated the dependence of electrotransfection efficiency (eTE) on binding of plasmid DNA (pDNA) to plasma membrane. Binding concentrates DNA molecules in the vicinity of the cell membrane, which should theoretically result in a greater number of DNA-membrane interactions during pulsed electric field, more internalized DNA, and ultimately, higher eTE values. We demonstrated that supplementing the electrotransfection buffer with divalent cations (Ca2+ and Mg2+) is an effective method of promoting pDNA adsorption to the cell membrane. This cation-mediated increase in DNA adsorption to the cellular membrane resulted in a consequent increase in eTE, up to a certain threshold concentration for each cation. To determine the timeframe for completion of pDNA internalization following pulse treatment, trypsin treatment was applied to cells at different timepoints after electrotransfection to strip off any residual, membrane-bound pDNA that had not been internalized. Trypsin treatment at 10 min post electrotransfection still resulted in a significant reduction in eTE, indicating that the time period for complete cellular uptake far exceeded the lifetime (~ 10 msec) of electric field-induced transient pores. The role of endocytosis was further probed by noting the effect on eTE when cells were treated with three endocytic inhibitors (chlorpromazine, genistein, dynasore) targeting different internalization…
Advisors/Committee Members: Yuan, Fan (advisor).
Subjects/Keywords: Biomedical engineering;
Electric Field;
Gene Delivery;
Transfection
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APA ·
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APA (6th Edition):
Wu, M. (2011). Implicated Role of Endocytosis in the Internalization and Intracellular Transport of Plasmid DNA During Electric Field-Mediated Gene Delivery
. (Thesis). Duke University. Retrieved from http://hdl.handle.net/10161/4994
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Wu, Mina. “Implicated Role of Endocytosis in the Internalization and Intracellular Transport of Plasmid DNA During Electric Field-Mediated Gene Delivery
.” 2011. Thesis, Duke University. Accessed January 22, 2020.
http://hdl.handle.net/10161/4994.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Wu, Mina. “Implicated Role of Endocytosis in the Internalization and Intracellular Transport of Plasmid DNA During Electric Field-Mediated Gene Delivery
.” 2011. Web. 22 Jan 2020.
Vancouver:
Wu M. Implicated Role of Endocytosis in the Internalization and Intracellular Transport of Plasmid DNA During Electric Field-Mediated Gene Delivery
. [Internet] [Thesis]. Duke University; 2011. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/10161/4994.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Wu M. Implicated Role of Endocytosis in the Internalization and Intracellular Transport of Plasmid DNA During Electric Field-Mediated Gene Delivery
. [Thesis]. Duke University; 2011. Available from: http://hdl.handle.net/10161/4994
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Hong Kong University of Science and Technology
16.
Gao, Wenhui.
Study of DNA-uptake mechanisms in electroporation.
Degree: 1998, Hong Kong University of Science and Technology
URL: http://repository.ust.hk/ir/Record/1783.1-3705
;
https://doi.org/10.14711/thesis-b589613
;
http://repository.ust.hk/ir/bitstream/1783.1-3705/1/th_redirect.html
► Electroporation is one of the most common methods used to transfer exogenous gene into most prokaryotic or eukaryotic cells. The detailed mechanisms of DNA-uptake by…
(more)
▼ Electroporation is one of the most common methods used to transfer exogenous gene into most prokaryotic or eukaryotic cells. The detailed mechanisms of DNA-uptake by cells, however, have not yet been clearly understood. In this thesis, we have summarized a series of experiment designed to examine the DAN- uptake mechanisms. In addition, cell treatments that can enhance the efficiency of electroporation are also discussed. The results show that after the application of electric field the additional treatment by using a recovery medium containing proper amounts of divalent ions can significantly increase the transfection efficiency in 3T6, Pt K2 and SK-N-SH cells. Several factors, including the strength of electric field, recovery time and concentrations of Ca++ or Mg++ were examined. The reason for increasing the transfection efficiency is believed to be mainly caused by promoting the entry of DNA through the cell membrane, rather than by enhancing the expression of the transferred gene. It has been suggested that DNA-uptake is a multiple- step process. (1) DNA may be driven into the cell by an electrophoretic force when the membrane pores were opened during the application of the electric field. (2) After the electric field was turned off, DNA may continue to diffuse into the cell before the membrane pores were closed. (3) Finally, DNA may be carried in the cell by endocytosis after the membrane pores were resealed. In this research, we have designed specific experiments to test each of these steps. Our results support steps (1) and (3), but not step (2). We found that the kinetics of membrane pore resealing depends strongly on the size of the transport molecules. Thus, uptake of large molecules such as FITC labeled dextran (70 kD) or DNA stopped almost within seconds after the electric field was turned off. In summary, we propose that the DNA-uptake mechanisms in electroporation involve a fast step of DNA entrance driven by the electric field, and a relatively slower process of DNA-uptake by endocytosis following the turn-off of the electric pulses. The experimental evidence will be discussed in detail in this thesis.
Subjects/Keywords: Electroporation
; DNA
; Transfection
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APA (6th Edition):
Gao, W. (1998). Study of DNA-uptake mechanisms in electroporation. (Thesis). Hong Kong University of Science and Technology. Retrieved from http://repository.ust.hk/ir/Record/1783.1-3705 ; https://doi.org/10.14711/thesis-b589613 ; http://repository.ust.hk/ir/bitstream/1783.1-3705/1/th_redirect.html
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Gao, Wenhui. “Study of DNA-uptake mechanisms in electroporation.” 1998. Thesis, Hong Kong University of Science and Technology. Accessed January 22, 2020.
http://repository.ust.hk/ir/Record/1783.1-3705 ; https://doi.org/10.14711/thesis-b589613 ; http://repository.ust.hk/ir/bitstream/1783.1-3705/1/th_redirect.html.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Gao, Wenhui. “Study of DNA-uptake mechanisms in electroporation.” 1998. Web. 22 Jan 2020.
Vancouver:
Gao W. Study of DNA-uptake mechanisms in electroporation. [Internet] [Thesis]. Hong Kong University of Science and Technology; 1998. [cited 2020 Jan 22].
Available from: http://repository.ust.hk/ir/Record/1783.1-3705 ; https://doi.org/10.14711/thesis-b589613 ; http://repository.ust.hk/ir/bitstream/1783.1-3705/1/th_redirect.html.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Gao W. Study of DNA-uptake mechanisms in electroporation. [Thesis]. Hong Kong University of Science and Technology; 1998. Available from: http://repository.ust.hk/ir/Record/1783.1-3705 ; https://doi.org/10.14711/thesis-b589613 ; http://repository.ust.hk/ir/bitstream/1783.1-3705/1/th_redirect.html
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Louisiana State University
17.
Wilson, Jessica A.
Determining gene copy number in transfected caprine fibroblast cells.
Degree: MS, Animal Sciences, 2009, Louisiana State University
URL: etd-04082009-113609
;
https://digitalcommons.lsu.edu/gradschool_theses/826
► Transgene expression in stably transgenic organisms is affected by many factors, including the copy number of the transgene in the genome, and by interactions between…
(more)
▼ Transgene expression in stably transgenic organisms is affected by many factors, including the copy number of the transgene in the genome, and by interactions between the transgene and flanking DNA sequences. Transgene copy number has also been shown to effect genetic stability in transgenic plants. Two commonly used methods for transfecting cells are liposome mediated transfection and electroporation. Little is known about the mean transgene copy number or variability of the copy number with these techniques. Quantitative PCR (Q-PCR) has been shown to be an effective method for determining transgene copy number. The objective of this study was to determine transgene copy number after liposome mediated transfection and electroporation. The mean transgene copy number and variability between individual integration events have been determined. Q-PCR conditions were optimized for primer annealing temperature and concentration when amplifying a region of the plasmid hEFGFP used for transfection. The quantitative nature of the Q-PCR reaction was confirmed by amplifying 10-fold dilutions of the plasmid and plotting the threshold cycle (CT) value against the log of the plasmid concentration. A correlation coefficient of 1.00 and a calculated PCR efficiency of 93.3% were obtained from this analysis. Caprine fibroblasts were transfected by electroporation or FuGENE® HD reagent with either a circular or linearized hEFGFP plasmid and plated at low density in medium containing Geneticin®. After 10 days of culture, single cell colonies were isolated and expanded. When cultures reached 1-2 million cells, genomic DNA was isolated. Transgene copy number was determined by amplifying genomic DNA from individual clones representing 1x105 cells with Q-PCR. Transgene copy number was calculated by comparing CT values to a standard curve. The transgene copy number for electroporation circular was 2.7 0.75 (n=32) and 1.3 0.65 (n=19) when using a linear DNA construct. FuGENE® HD using a circular plasmid construct generated a gene copy number of 0.5 0.11 (n=14) and 0.64 0.13 (n=16) for the linear plasmid construct. There were significant differences when comparing electroporation circular to all other treatments, however, there were no differences when comparing electroporation linear, FuGENE® HD circular and FuGENE® HD linear to each other.
Subjects/Keywords: Transgenic; Transgene copy number; Transfection; Quantitative PCR; Plasmid; Liposome mediated transfection; FuGENE HD; Electroporation; Caprine
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APA (6th Edition):
Wilson, J. A. (2009). Determining gene copy number in transfected caprine fibroblast cells. (Masters Thesis). Louisiana State University. Retrieved from etd-04082009-113609 ; https://digitalcommons.lsu.edu/gradschool_theses/826
Chicago Manual of Style (16th Edition):
Wilson, Jessica A. “Determining gene copy number in transfected caprine fibroblast cells.” 2009. Masters Thesis, Louisiana State University. Accessed January 22, 2020.
etd-04082009-113609 ; https://digitalcommons.lsu.edu/gradschool_theses/826.
MLA Handbook (7th Edition):
Wilson, Jessica A. “Determining gene copy number in transfected caprine fibroblast cells.” 2009. Web. 22 Jan 2020.
Vancouver:
Wilson JA. Determining gene copy number in transfected caprine fibroblast cells. [Internet] [Masters thesis]. Louisiana State University; 2009. [cited 2020 Jan 22].
Available from: etd-04082009-113609 ; https://digitalcommons.lsu.edu/gradschool_theses/826.
Council of Science Editors:
Wilson JA. Determining gene copy number in transfected caprine fibroblast cells. [Masters Thesis]. Louisiana State University; 2009. Available from: etd-04082009-113609 ; https://digitalcommons.lsu.edu/gradschool_theses/826
18.
Pérard, Matthieu.
Mise au point d’un modèle tridimensionnel de culture d’odontoblastes. Application à l’évaluation in vitro de biomatériaux. : Development of a three-dimensional model of odontoblast culture. Application to an in vitro evaluation of biomaterials.
Degree: Docteur es, Biologie et sciences de la santé, 2015, Rennes 1
URL: http://www.theses.fr/2015REN1B001
► L’objectif de ce travail était de mettre au point un modèle de culture cellulaire, afin d’étudier in vitro l’incidence sur la physiologie des cellules pulpaires…
(more)
▼ L’objectif de ce travail était de mettre au point un modèle de culture cellulaire, afin d’étudier in vitro l’incidence sur la physiologie des cellules pulpaires et en particulier de l’odontoblaste, de biomatériaux utilisés pour traiter les effractions de pulpes dentaires. Ce modèle repose sur l’utilisation de culture sphéroïdes dont la conformation spatiale reproduit plus fidèlement l’environnement in vivo que les cultures bidimensionnelles. Après avoir élaboré le modèle sphéroïde à partir de lignées murines, des expérimentations visant à déterminer la cytotoxicité des matériaux ont été effectuées. Leurs capacités à induire la biominéralisation ont également été évaluées. La dernière partie de ce travail avait pour objectif d’immortaliser des primocultures de cellules pulpaires humaines en transfectant les gènes SV40 et hTERT, afin d’établir une lignée cellulaire
The aim of this study was to develop a cell culture model to assess in vitro the effects on the physiology of pulp cells, in particular the odontoblasts, of biomaterials used to treat dental pulp exposures. This model is based on the use of spheroid culture whose spatial configuration reproduces the in vivo environment more faithfully than do two-dimensional cultures. After developing the spheroid model from mouse lines, experiments to determine the cytotoxicity of the materials were conducted. Their ability to induce bio-mineralisation was also assessed. The last part of this work aimed to immortalise primo-cultures of human pulp cells by transfecting hTERT and SV40 genes, in order to establish a new cell line
Advisors/Committee Members: Pérez, Fabienne (thesis director).
Subjects/Keywords: Sphéroïdes; Cellules pulpaires; Minéralisation; Coiffage pulpaire; Transfection; Spheroids; Pulp cells; Mineralisation; Pulp capping; Transfection
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APA (6th Edition):
Pérard, M. (2015). Mise au point d’un modèle tridimensionnel de culture d’odontoblastes. Application à l’évaluation in vitro de biomatériaux. : Development of a three-dimensional model of odontoblast culture. Application to an in vitro evaluation of biomaterials. (Doctoral Dissertation). Rennes 1. Retrieved from http://www.theses.fr/2015REN1B001
Chicago Manual of Style (16th Edition):
Pérard, Matthieu. “Mise au point d’un modèle tridimensionnel de culture d’odontoblastes. Application à l’évaluation in vitro de biomatériaux. : Development of a three-dimensional model of odontoblast culture. Application to an in vitro evaluation of biomaterials.” 2015. Doctoral Dissertation, Rennes 1. Accessed January 22, 2020.
http://www.theses.fr/2015REN1B001.
MLA Handbook (7th Edition):
Pérard, Matthieu. “Mise au point d’un modèle tridimensionnel de culture d’odontoblastes. Application à l’évaluation in vitro de biomatériaux. : Development of a three-dimensional model of odontoblast culture. Application to an in vitro evaluation of biomaterials.” 2015. Web. 22 Jan 2020.
Vancouver:
Pérard M. Mise au point d’un modèle tridimensionnel de culture d’odontoblastes. Application à l’évaluation in vitro de biomatériaux. : Development of a three-dimensional model of odontoblast culture. Application to an in vitro evaluation of biomaterials. [Internet] [Doctoral dissertation]. Rennes 1; 2015. [cited 2020 Jan 22].
Available from: http://www.theses.fr/2015REN1B001.
Council of Science Editors:
Pérard M. Mise au point d’un modèle tridimensionnel de culture d’odontoblastes. Application à l’évaluation in vitro de biomatériaux. : Development of a three-dimensional model of odontoblast culture. Application to an in vitro evaluation of biomaterials. [Doctoral Dissertation]. Rennes 1; 2015. Available from: http://www.theses.fr/2015REN1B001
Brno University of Technology
19.
Pošustová, Veronika.
Transfekce somatických buněk pomocí magnetických nanočástic a jejich post-transfekční lokalizace
.
Degree: 2018, Brno University of Technology
URL: http://hdl.handle.net/11012/82553
► Tato bakalářská práce je zaměřena na experiment založený na magnetických nanočástic SPIO pro transfekci 3T3 buněk. Povrch nanočástic byl modifikován pomocí PEI a Chitosanu za…
(more)
▼ Tato bakalářská práce je zaměřena na experiment založený na magnetických nanočástic SPIO pro transfekci 3T3 buněk. Povrch nanočástic byl modifikován pomocí PEI a Chitosanu za účelem optimalizace této metody. Transfekční experiment byl proveden dvanáct krát, následně byla nasnímaná data zpracována a statisticky vyhodnocena v programovacím prostředí Matlab R2016a a určena transfekční efektivita. Teoretická část je zaměřena na magnetické nanočástice a jejich využití v medicíně, dále na transfekci a její metody a také konkrétně na transfekční metodu založenou na magnetických nanočásticích.; This bachelor thesis is focused on an experiment based on magnetic particles SPIO for
transfection of 3T3 cells. Nanoparticle surface was modified with PEI and Chitosan to optimize this method. The
transfection experiment was performed twelve times, then the scanned data was processed and statistically analyzed in Matlab R2016a to determine the
transfection efficiency. The theoretical part is focused on magnetic nanoparticles and their application in medicine, on
transfection and in last part on a
transfection of cells using magnetic nanoparticles.
Advisors/Committee Members: Svoboda, Ondřej (advisor).
Subjects/Keywords: Nanočástice;
magnetické nanočástice;
transfekce;
transfekční efektivita;
Nanoparticles;
magnetic nanoparticles;
transfection;
transfection efficiency
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APA (6th Edition):
Pošustová, V. (2018). Transfekce somatických buněk pomocí magnetických nanočástic a jejich post-transfekční lokalizace
. (Thesis). Brno University of Technology. Retrieved from http://hdl.handle.net/11012/82553
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Pošustová, Veronika. “Transfekce somatických buněk pomocí magnetických nanočástic a jejich post-transfekční lokalizace
.” 2018. Thesis, Brno University of Technology. Accessed January 22, 2020.
http://hdl.handle.net/11012/82553.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Pošustová, Veronika. “Transfekce somatických buněk pomocí magnetických nanočástic a jejich post-transfekční lokalizace
.” 2018. Web. 22 Jan 2020.
Vancouver:
Pošustová V. Transfekce somatických buněk pomocí magnetických nanočástic a jejich post-transfekční lokalizace
. [Internet] [Thesis]. Brno University of Technology; 2018. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/11012/82553.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Pošustová V. Transfekce somatických buněk pomocí magnetických nanočástic a jejich post-transfekční lokalizace
. [Thesis]. Brno University of Technology; 2018. Available from: http://hdl.handle.net/11012/82553
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
20.
Raisin, Sophie.
Supports biomimétiques actifs pour la différenciation de cellules souches mésenchymateuses : application à la régénération du cartilage : Active biomimetic supports for mesenchymal stem cells : application to cartilage regeneration.
Degree: Docteur es, Chimie et physico-chimie des matériaux, 2016, Montpellier
URL: http://www.theses.fr/2016MONTT232
► La conception de biomatériaux actifs est actuellement encouragée par le manque de thérapies régénératives efficaces pour des tissus endommagés présentant une faible capacité d’autoréparation. Les…
(more)
▼ La conception de biomatériaux actifs est actuellement encouragée par le manque de thérapies régénératives efficaces pour des tissus endommagés présentant une faible capacité d’autoréparation. Les progrès récents concernant les techniques de préparation de matériaux structurés (électrospinning, microfluidique) ainsi que la découverte du fort potentiel régénératif des cellules souches ont suscité un regain d’intérêt pour des projets collaboratifs à l’interface entre biologie et sciences des matériaux. Une approche prometteuse de régénération tissulaire repose donc sur la combinaison de cellules souches et de biomatériaux implantables. Des biomatériaux innovants, injectables et servants à la fois de support aux cellules et de réservoir de molécules actives telles que des protéines ou des agents de thérapie génique (Matrice Génétiquement Activée) ont été développés. Se plaçant plus particulièrement dans le contexte de l’ingénierie du cartilage, ce travail a pour objectif de développer une stratégie complémentaire concernant l’orientation de la différenciation de cellules souches mésenchymateuses (CSM) grâce au mécanisme d’interférence ARN.La principale difficulté rencontrée lors de l’utilisation d’acides nucléiques pour induire la différenciation des CSM reste leur faible capacité à traverser les membranes cellulaires, due à leur nature hydrophile et leur charge négative. De plus, les acides nucléiques sont dégradés très facilement par les nucléases extracellulaires, ce qui rend nécessaire l’utilisation d’un vecteur. Les vecteurs non-viraux sont d’excellents candidats pour des applications in vivo en raison de leur faible coût de production et leur faible immunogénicité. Toutefois, la plupart des systèmes de vectorisation trouvés dans la littérature présentent un manque de reproductibilité associé à une cytotoxicité vis-à-vis des cellules primaires. Nous souhaitions donc développer un système de
transfection synthétique à la fois efficace et biocompatible. Pour cela, nous nous sommes basés sur les résultats encourageants concernant l’utilisation des micelles de complexes polyioniques (PIC) pour la
transfection des cellules dendritiques. Ces micelles sont formées par complexation de deux polyélectrolytes : un copolymère à blocs double-hydrophiles (CBDH) avec un bloc anionique et un homopolymère cationique. Dans ce travail, nous avons évalué le polyoxyde d’éthylène – b – polyacide méthacrylique en tant que CBDH et la poly-L-lysine ou le polyéthylènimine en tant que polycation. L’influence des caractéristiques des composantes (asymétrie du CBDH, nature du polycation, taille des blocs, ratio de charges…) sur les propriétés physico-chimiques des micelles formées (taille, charge de surface) a d’abord été étudiée. Puis, la possibilité de complexation d’un siRNA au sein des micelles ainsi que leur stabilité en conditions physiologiques ont été évaluées. La formulation des micelles a été conçue pour permettre une dissociation des objets à un pH comparable à celui des endosomes ; ceci a été vérifié par diffusion dynamique de la…
Advisors/Committee Members: Belamie, Emmanuel (thesis director).
Subjects/Keywords: Matériaux biofonctionnels; Cellules souches; Nanoparticules; Transfection; SiRNA; Matériaux biomimétiques; Biofunctional materials; Stem cells; Nanoparticles; Transfection; SiRNA; Biomimetic materials
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APA (6th Edition):
Raisin, S. (2016). Supports biomimétiques actifs pour la différenciation de cellules souches mésenchymateuses : application à la régénération du cartilage : Active biomimetic supports for mesenchymal stem cells : application to cartilage regeneration. (Doctoral Dissertation). Montpellier. Retrieved from http://www.theses.fr/2016MONTT232
Chicago Manual of Style (16th Edition):
Raisin, Sophie. “Supports biomimétiques actifs pour la différenciation de cellules souches mésenchymateuses : application à la régénération du cartilage : Active biomimetic supports for mesenchymal stem cells : application to cartilage regeneration.” 2016. Doctoral Dissertation, Montpellier. Accessed January 22, 2020.
http://www.theses.fr/2016MONTT232.
MLA Handbook (7th Edition):
Raisin, Sophie. “Supports biomimétiques actifs pour la différenciation de cellules souches mésenchymateuses : application à la régénération du cartilage : Active biomimetic supports for mesenchymal stem cells : application to cartilage regeneration.” 2016. Web. 22 Jan 2020.
Vancouver:
Raisin S. Supports biomimétiques actifs pour la différenciation de cellules souches mésenchymateuses : application à la régénération du cartilage : Active biomimetic supports for mesenchymal stem cells : application to cartilage regeneration. [Internet] [Doctoral dissertation]. Montpellier; 2016. [cited 2020 Jan 22].
Available from: http://www.theses.fr/2016MONTT232.
Council of Science Editors:
Raisin S. Supports biomimétiques actifs pour la différenciation de cellules souches mésenchymateuses : application à la régénération du cartilage : Active biomimetic supports for mesenchymal stem cells : application to cartilage regeneration. [Doctoral Dissertation]. Montpellier; 2016. Available from: http://www.theses.fr/2016MONTT232
21.
Andrianaivo, Fanjambolatiana.
Contribution à l'étude de la transfection hydrodynamique : effet de l'injection hydrodynamique sur l'endocytose par le foie.
Degree: 2004, DIAL (Belgium)
URL: http://hdl.handle.net/2078.2/4243
► Notre travail concerne la transfection génique. Celle-ci nécessite l'envoi au noyau cellulaire d'ADN exogène, c'est-à-dire, d'une grosse molécule hydrophile, qui, normalement ne peut pénétrer dans…
(more)
▼ Notre travail concerne la transfection génique. Celle-ci nécessite l'envoi au noyau cellulaire d'ADN exogène, c'est-à-dire, d'une grosse molécule hydrophile, qui, normalement ne peut pénétrer dans la cellule que par endocytose. Or, un tel processus conduit habituellement à une dégradation de la molécule, en grande partie dans les lysosomes. Dans notre introduction, après un bref rappel sur l'endocytose, nous présenterons les moyens qui permettent à l'ADN exogène d'échapper à une hydrolyse intracellulaire et d'atteindre le noyau sous une forme fonctionnelle.
Les résultats que nous avons présentés nous apportent des informations sur l'influence de l'injection hydrodynamique sur l'endocytose par le foie. Rappelons que notre travail avait deux buts principaux : contribuer à l'élucidation du mécanisme de la transfection hydrodynamique et rechercher si l'injection hydrodynamique pouvait présenter des avantages pour introduire des protéines dans les cellules hépatiques.
(DOCMED00) – FUNDP, 2004
Advisors/Committee Members: FUNDP - 2851 - Laboratoire de Physiologie Cellulaire, Michel JADOT, WATTIAUX-DE CONINCK, Simone, DE BACKER , Olivier, DEVOS, Pierre, MERTENS-STRIJTHAGEN, Jeannine, WIBO, Maurice.
Subjects/Keywords: ADN; endocytose; transfection hydrodynamique; lysosomes; injection hydrodynamique; Transfection génique
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APA (6th Edition):
Andrianaivo, F. (2004). Contribution à l'étude de la transfection hydrodynamique : effet de l'injection hydrodynamique sur l'endocytose par le foie. (Thesis). DIAL (Belgium). Retrieved from http://hdl.handle.net/2078.2/4243
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Andrianaivo, Fanjambolatiana. “Contribution à l'étude de la transfection hydrodynamique : effet de l'injection hydrodynamique sur l'endocytose par le foie.” 2004. Thesis, DIAL (Belgium). Accessed January 22, 2020.
http://hdl.handle.net/2078.2/4243.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Andrianaivo, Fanjambolatiana. “Contribution à l'étude de la transfection hydrodynamique : effet de l'injection hydrodynamique sur l'endocytose par le foie.” 2004. Web. 22 Jan 2020.
Vancouver:
Andrianaivo F. Contribution à l'étude de la transfection hydrodynamique : effet de l'injection hydrodynamique sur l'endocytose par le foie. [Internet] [Thesis]. DIAL (Belgium); 2004. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/2078.2/4243.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Andrianaivo F. Contribution à l'étude de la transfection hydrodynamique : effet de l'injection hydrodynamique sur l'endocytose par le foie. [Thesis]. DIAL (Belgium); 2004. Available from: http://hdl.handle.net/2078.2/4243
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Florida
22.
Wang, Xinyi.
Development of a Transfection System for Genetic Manipulation of Babesia Bovis.
Degree: PhD, Medical Sciences - Immunology and Microbiology (IDP), 2010, University of Florida
URL: http://ufdc.ufl.edu/UFE0041150
► Babesia bovis is an intraerythrocytic parasite which establishes and maintains a persistent infection in its bovine host by at least two mechanisms: cytoadhesion and antigenic…
(more)
▼ Babesia bovis is an intraerythrocytic parasite which establishes and maintains a persistent infection in its bovine host by at least two mechanisms: cytoadhesion and antigenic variation. In the past few years, a
transfection system has been initiated in B. bovis, providing a useful genetic tool for studying the biology of the parasite. Here I present further development and extension of
transfection methodology for B. bovis, as a technical basis for targeted genetic manipulation of this parasite. I have determined that B. bovis parasites are adequately susceptible to puromycin, blasticidin-S and pyrimethamine to provide for positive selection, using introduction of an exogenous gene imparting resistance, making stable transformation feasible. In developing this approach, I have established a transient
transfection system employing firefly luciferase expression to compare promoter activities, and have demonstrated that the IGr s of the LAT and two
transcriptionally silent ves donor gene pairs all display very strong promoter activities, at levels comparable to promoter from the housekeeping gene calmodulin. Taking advantage of this technology to gain insights into the gene regulation in the unique intergenic region of LAT, I have also identified a cluster of promoters within the 434 bp sequences as well as possible enhancing activities embedded in the flanking exonic or intronic sequences. Interestingly, one inverted intron has resulted in significant reduction in promoter activities, suggesting possible DNA binding motifs in that region. Besides, transient expression of enhanced green fluorescent protein whose expression is driven by a ves promoter has revealed there is no strict ves promoter counting in the parasite when only promoter sequences are present. Thus, ves gene expression from the LAT was unaffected while the episomal vector expressing EGFP was replicating and transcriptionally active extrachromosomally. Stable
transfection of the parasite has also been achieved, revealing a non-targeted integration event with stable expression of gfp-bsd protein in the transfectants, which suggests it may be difficult to reproducibly achieve the specific genetic alterations desired. These observations provide key background knowledge and reagents for deliberate genetic manipulation of B. bovis. ( en )
Advisors/Committee Members: Allred, David R. (committee chair), Mergia, Ayalew (committee member), Barbet, Anthony F. (committee member), Jin, Shouguang (committee member), Bloom, Linda B. (committee member).
Subjects/Keywords: Antigenic variation; DNA; Erythrocytes; Fireflies; Genes; Intergenic DNA; Introns; Parasites; Plasmids; Transfection; babesia, bovis, lat, promoter, transfection, ves
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APA (6th Edition):
Wang, X. (2010). Development of a Transfection System for Genetic Manipulation of Babesia Bovis. (Doctoral Dissertation). University of Florida. Retrieved from http://ufdc.ufl.edu/UFE0041150
Chicago Manual of Style (16th Edition):
Wang, Xinyi. “Development of a Transfection System for Genetic Manipulation of Babesia Bovis.” 2010. Doctoral Dissertation, University of Florida. Accessed January 22, 2020.
http://ufdc.ufl.edu/UFE0041150.
MLA Handbook (7th Edition):
Wang, Xinyi. “Development of a Transfection System for Genetic Manipulation of Babesia Bovis.” 2010. Web. 22 Jan 2020.
Vancouver:
Wang X. Development of a Transfection System for Genetic Manipulation of Babesia Bovis. [Internet] [Doctoral dissertation]. University of Florida; 2010. [cited 2020 Jan 22].
Available from: http://ufdc.ufl.edu/UFE0041150.
Council of Science Editors:
Wang X. Development of a Transfection System for Genetic Manipulation of Babesia Bovis. [Doctoral Dissertation]. University of Florida; 2010. Available from: http://ufdc.ufl.edu/UFE0041150
23.
Sabraoui, Abbas.
Régulation de la cavitation acoustique appliquée à la transfection cellulaire : Feedback loop process to control acoustic cavitation : application to cell transfection.
Degree: Docteur es, Acoustique, 2012, Université Claude Bernard – Lyon I
URL: http://www.theses.fr/2012LYO10007
► Le travail présenté ici porte sur l’étude et le contrôle de la cavitation acoustique dans le but de développer un système de sonoporation efficace pour…
(more)
▼ Le travail présenté ici porte sur l’étude et le contrôle de la cavitation acoustique dans le but de développer un système de sonoporation efficace pour les cellules en suspension et les cellules adhérentes. Le manuscrit est composé de trois chapitres. Tout d’abord, une revue de la littérature sur les différentes techniques physiques utilisées en transfection cellulaire, et plus particulièrement la sonoporation. Il a été démontré que le principal mécanisme de la sonoporation est étroitement lié au phénomène de cavitation acoustique. Un contrôle de ce phénomène aléatoire apparaît alors intéressant afin d’augmenter le taux de transfection tout en gardant une forte viabilité cellulaire. Dans le second chapitre, un système de régulation de cavitation ultrasonore basé sur un indice acoustique de cavitation a été étudié. Cet indice, est basé sur la mesure de bruit large bande émis lors de l’implosion des bulles de cavitation. Les avantages d’un tel système sont : un suivi en temps réel du niveau de cavitation durant l’irradiation, des informations quasi-instantanées sur les composantes spectrales caractéristiques de la cavitation, une meilleure reproductibilité et stabilité du niveau de cavitation surtout pour les intensités modérés. Dans le troisième chapitre, pour comprendre les mécanismes de la sonoporation, un deuxième système de cavitation contrôlé a été conçu dans le but de permettre une visualisation du milieu en cours d’insonification. Ce nouveau dispositif est adapté à un fonctionnement sous microscope photonique à transmission et à fluorescence. Des essais de transfection de siRNAs, sur les cellules en suspension (RL du lymphome folliculaire) et sur les cellules adhérentes (cancer du sein ; MDA-MB 231) ont permis de valider in vitro l’efficacité de ce système en atteignant un taux de 40 % de transfection pour ces deux types de cellules, avec un très faible taux de mortalité (< 10 %)
The aim of the present work, which is based on the study and the control of acoustic cavitation, is to develop an efficient sonoporation system to transfect the cells in suspension and the adherent cells. The manuscript is composed of three chapters. The first one takes a glance on the state of art of different physical techniques used in cells transfection, and more precisely on sonoporation. It has been shown that the principal mechanism of sonoporation is closely linked to acoustic cavitation. Thus, a control of this random phenomenon is important to increase the rate of transfection while keeping strong cell viability. In the second chapter, a regulated cavitation generator based on an acoustic index was studied. This index is based on the measure of broad band noise emitted during the implosion of the cavitation bubbles. The advantage of such a system is: a control in real time of the level cavitation during sonication, leading to a better reproducibility and stability of the cavitation level, especially for the moderate intensities. In the third chapter, in order further study the sonoporation mechanisms, a second regulated…
Advisors/Committee Members: Béra, Jean-Christophe (thesis director), Mestas, Jean-Louis (thesis director).
Subjects/Keywords: Acoustique; Cavitation; Ultrasons; Sonoporation; Seuil de cavitation; Régulation; Transfection; Acoustics; Cavitation; Ultrasound; Sonoporation; Cavitation threshold; Regulation; Transfection; 620.2
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APA (6th Edition):
Sabraoui, A. (2012). Régulation de la cavitation acoustique appliquée à la transfection cellulaire : Feedback loop process to control acoustic cavitation : application to cell transfection. (Doctoral Dissertation). Université Claude Bernard – Lyon I. Retrieved from http://www.theses.fr/2012LYO10007
Chicago Manual of Style (16th Edition):
Sabraoui, Abbas. “Régulation de la cavitation acoustique appliquée à la transfection cellulaire : Feedback loop process to control acoustic cavitation : application to cell transfection.” 2012. Doctoral Dissertation, Université Claude Bernard – Lyon I. Accessed January 22, 2020.
http://www.theses.fr/2012LYO10007.
MLA Handbook (7th Edition):
Sabraoui, Abbas. “Régulation de la cavitation acoustique appliquée à la transfection cellulaire : Feedback loop process to control acoustic cavitation : application to cell transfection.” 2012. Web. 22 Jan 2020.
Vancouver:
Sabraoui A. Régulation de la cavitation acoustique appliquée à la transfection cellulaire : Feedback loop process to control acoustic cavitation : application to cell transfection. [Internet] [Doctoral dissertation]. Université Claude Bernard – Lyon I; 2012. [cited 2020 Jan 22].
Available from: http://www.theses.fr/2012LYO10007.
Council of Science Editors:
Sabraoui A. Régulation de la cavitation acoustique appliquée à la transfection cellulaire : Feedback loop process to control acoustic cavitation : application to cell transfection. [Doctoral Dissertation]. Université Claude Bernard – Lyon I; 2012. Available from: http://www.theses.fr/2012LYO10007
24.
Morin-Picardat, Emmanuelle.
Etudes d'auto-assemblages polydiacétylèniques et applications biologiques : Polydiacetylenic self-assemblies and biological applications.
Degree: Docteur es, Chimie, 2012, Université de Strasbourg
URL: http://www.theses.fr/2012STRAF063
► La dualité hydrophobe/hydrophile des molécules amphiphiles est à l’origine de leur autoassemblage en solution, sous forme de nombreuses structures supramoléculaires, telles que les micelles. Ce…
(more)
▼ La dualité hydrophobe/hydrophile des molécules amphiphiles est à l’origine de leur autoassemblage en solution, sous forme de nombreuses structures supramoléculaires, telles que les micelles. Ce travail de thèse présente la formation, la caractérisation et l’étude de nouvelles micelles diacétylèniques photopolymérisables. Une première partie décrit ainsi lasynthèse de nouvelles micelles cationiques et l’étude de leur utilisation en tant qu’agent de transfert de gènes. Dans une seconde partie, nos travaux présentent l’étude de micelles polydiacétylèniques, porteuses de têtes polaires octaéthylèneglycol, comme potentiel système de délivrance de médicament. Les propriétés d’encapsulation de ces micelles ont été évaluées en présence d’un dérivé fullerène fluorescent. Puis, l’incorporation d’une sonde membranaire dans leur couronne lipophile a permis de réaliser une étude de leurs propriétés de délivrance in vitro. Une étude préliminaire de leur biodistribution in vivo a également été réalisée par tomographie à émission monophotonique grâce à la chélation d’un isotoperadioactif sur la surface des micelles. Enfin une dernière partie présente l’analyse de deux nouveaux auto-assemblages tubulaires obtenus au cours de nos travaux.
The hydrophobic/hydrophilic duality of amphiphilic molecules is at the origin of their selfassembly in solution, into many supramolecular structures such as micelles. This thesis presents the formation, characterization and study of new photopolymerizable diacetylenic micelles. The first part describes the synthesis of new cationic micelles and the study of theiruse as gene transfer agent. In the second part, our work presents the study of polydiacetylene micelles, bearing polar heads octaétylèneglycol as potential drug delivery system. The encapsulation properties of these micelles were first evaluated in the presence of a fluorescent fullerene derivative. Then, the incorporation of a membrane probe in their lipophilic shell has allowed a study of their in vitro delivery properties. Finally a study of their in vivo biodistribution was also carried out by single photon emission tomography through the chelation of a radioactive isotope on micelle surface. Finally the last part presents theanalysis of two new tubularself-assemblies, obtained during our work.
Advisors/Committee Members: Remy, Jean-Serge (thesis director), Wagner, Alain (thesis director).
Subjects/Keywords: Amphiphiles; Micelles; Polydiacétylènes; Transfection; Délivrance de médicaments; Auto-assemblages; Amphiphiles; Micelles; Polydiacetylenes; Transfection; Drug delivery; Self-assembling; 547.8
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APA (6th Edition):
Morin-Picardat, E. (2012). Etudes d'auto-assemblages polydiacétylèniques et applications biologiques : Polydiacetylenic self-assemblies and biological applications. (Doctoral Dissertation). Université de Strasbourg. Retrieved from http://www.theses.fr/2012STRAF063
Chicago Manual of Style (16th Edition):
Morin-Picardat, Emmanuelle. “Etudes d'auto-assemblages polydiacétylèniques et applications biologiques : Polydiacetylenic self-assemblies and biological applications.” 2012. Doctoral Dissertation, Université de Strasbourg. Accessed January 22, 2020.
http://www.theses.fr/2012STRAF063.
MLA Handbook (7th Edition):
Morin-Picardat, Emmanuelle. “Etudes d'auto-assemblages polydiacétylèniques et applications biologiques : Polydiacetylenic self-assemblies and biological applications.” 2012. Web. 22 Jan 2020.
Vancouver:
Morin-Picardat E. Etudes d'auto-assemblages polydiacétylèniques et applications biologiques : Polydiacetylenic self-assemblies and biological applications. [Internet] [Doctoral dissertation]. Université de Strasbourg; 2012. [cited 2020 Jan 22].
Available from: http://www.theses.fr/2012STRAF063.
Council of Science Editors:
Morin-Picardat E. Etudes d'auto-assemblages polydiacétylèniques et applications biologiques : Polydiacetylenic self-assemblies and biological applications. [Doctoral Dissertation]. Université de Strasbourg; 2012. Available from: http://www.theses.fr/2012STRAF063
25.
Sigwalt, David.
Conception, synthèse et étude de dérivés de C60 fonctionnalisés : applications biologiques et développement méthodologique : Design, synthesis and study of functionalized C60 : biological applications and methodological development.
Degree: Docteur es, Chimie, 2013, Université de Strasbourg
URL: http://www.theses.fr/2013STRAF010
► Notre équipe a récemment développé une méthode polyvalente permettant de préparer des dérivés complexes de C60 hexa-adduits fonctionnalisés. Cette méthodologie permet d’obtenir des produits aux…
(more)
▼ Notre équipe a récemment développé une méthode polyvalente permettant de préparer des dérivés complexes de C60 hexa-adduits fonctionnalisés. Cette méthodologie permet d’obtenir des produits aux caractéristiques originales. Le C60 central agit comme un support central peu réactif, autour duquel des fonctionnalités sont réparties dans un espace octaédrique parfaitement défini. La première partie de ce travail de thèse a consisté à exploiter cette méthodologie pour créer des C60 hexa-adduits polycationiques aux propriétés de transfection remarquables. Dans un second temps, les dendrons polyamines synthétisés ont été mis à profit pour créer des structures supramoléculaires de C60 hexa-adduits, sous forme micellaire. Par la suite, l’étude de ces assemblages a orienté nos investigations vers l’élaboration de dérivés de C60 hexa-adduits mannosylés multivalents résultant d’un assemblage supramoléculaire, dont leurs possibles applications biologiques sont actuellement à l’étude. En parallèle une synthèse covalente a permis d’obtenir un «équivalent dendritique» de C60 hexa-adduit multimannosylé. Partant du constat que notre méthodologie est efficace principalement pour des dérivés de C60 hexa-adduits qui ont une régio-sélectivité particulière, la dernière partie a été consacrée au développement de nouvelles voies de synthèses qui pourront permettre de créer des dérivés de C60 avec un contrôle régio-sélectif original.
Our team has recently built a polyvalent method that gives complex functionalized C60 hexa-adducts. This methodology permits to obtain products with original features. C60 acts as an inert scaffold, around which functionnalities are distributed in a well-defined octahedral space. The first part of this thesis describes exploitation of this methodology to create polycationics C60 hexa-adducts, which have shown remarkable gene delivery capabilities. Next, the synthetized polyamine dendrons were used to build supramolecular structure of C60 hexa-adducts, as micellar forms. The study of these self-assembled structures has guided us to design micelles of mannosylated C60 hexa-adducts, which biological applications are under investigation. In parallel a covalent synthesis has furnished a dendrimers-like multimannosylated C60 hexa-adduct. Based on the observation that our methodology to create C60 hexa-adduct is efficient only for a specific regio-selectivity, the last part of this thesis was devoted to the development of new synthetic routes to obtain C60 derivates with an original regio-selective control.
Advisors/Committee Members: Nierengarten, Jean-François (thesis director), Remy, Jean-Serge (thesis director).
Subjects/Keywords: C60; Dendrimer; Amphiphile; Micelles; Polydiacétylènes; Transfection; Auto-assemblages; C60; Dendrimers; Amphiphiles; Micelles; Polydiacetylenes; Transfection; Self-assembling; 547.2
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APA ·
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APA (6th Edition):
Sigwalt, D. (2013). Conception, synthèse et étude de dérivés de C60 fonctionnalisés : applications biologiques et développement méthodologique : Design, synthesis and study of functionalized C60 : biological applications and methodological development. (Doctoral Dissertation). Université de Strasbourg. Retrieved from http://www.theses.fr/2013STRAF010
Chicago Manual of Style (16th Edition):
Sigwalt, David. “Conception, synthèse et étude de dérivés de C60 fonctionnalisés : applications biologiques et développement méthodologique : Design, synthesis and study of functionalized C60 : biological applications and methodological development.” 2013. Doctoral Dissertation, Université de Strasbourg. Accessed January 22, 2020.
http://www.theses.fr/2013STRAF010.
MLA Handbook (7th Edition):
Sigwalt, David. “Conception, synthèse et étude de dérivés de C60 fonctionnalisés : applications biologiques et développement méthodologique : Design, synthesis and study of functionalized C60 : biological applications and methodological development.” 2013. Web. 22 Jan 2020.
Vancouver:
Sigwalt D. Conception, synthèse et étude de dérivés de C60 fonctionnalisés : applications biologiques et développement méthodologique : Design, synthesis and study of functionalized C60 : biological applications and methodological development. [Internet] [Doctoral dissertation]. Université de Strasbourg; 2013. [cited 2020 Jan 22].
Available from: http://www.theses.fr/2013STRAF010.
Council of Science Editors:
Sigwalt D. Conception, synthèse et étude de dérivés de C60 fonctionnalisés : applications biologiques et développement méthodologique : Design, synthesis and study of functionalized C60 : biological applications and methodological development. [Doctoral Dissertation]. Université de Strasbourg; 2013. Available from: http://www.theses.fr/2013STRAF010
Brno University of Technology
26.
Bílek, Ondřej.
Využití magnetických nanočástic pro transfekci plasmidu do HEK293 buněk
.
Degree: 2015, Brno University of Technology
URL: http://hdl.handle.net/11012/40709
► Tato bakalářská práce se zabývá využitím magnetických nanočástic, SPIO nanočástic a nanočástic komerčního produktu MATra pro transfekci plasmidu ASAP do HEK293 buněk a následnou optimalizací…
(more)
▼ Tato bakalářská práce se zabývá využitím magnetických nanočástic, SPIO nanočástic a nanočástic komerčního produktu MATra pro transfekci plasmidu ASAP do HEK293 buněk a následnou optimalizací tohoto procesu. K experimentální části je přiložena teoretická část shrnující poznatky z oblasti syntézy magnetických nanočástic, jejich využití v biomedicíně, teorii zabývající se možnostmi vložení cizorodé DNA či plasmidu do buňky a způsoby vyhodnocování úspěšnosti těchto experimentů.; This bachelor’s thesis deals with using of magnetic nanoparticles, SPIO nanoparticles and nanoparticles of commercial product MATra for
transfection of ASAP plasmid into HEK293 cells and subsequent optimization of the process. The experimental part is enclosed with theorethical part that summarizes findings from the fields of synthesis of magnetic nanoparticles and their biomedical applications, theory dealing with possibilities of the insertion of foreign DNA or plasmids into the cell and of its efficiency evaluation.
Advisors/Committee Members: Fohlerová, Zdenka (advisor).
Subjects/Keywords: Magnetické nanočástice;
SPIO nanočástice;
MATra;
transfekce;
DNA;
účinnost transfekce;
Magnetic nanoparticles;
SPIO nanoparticles;
MATra;
transfection;
DNA;
transfection efficiency
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APA ·
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APA (6th Edition):
Bílek, O. (2015). Využití magnetických nanočástic pro transfekci plasmidu do HEK293 buněk
. (Thesis). Brno University of Technology. Retrieved from http://hdl.handle.net/11012/40709
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Bílek, Ondřej. “Využití magnetických nanočástic pro transfekci plasmidu do HEK293 buněk
.” 2015. Thesis, Brno University of Technology. Accessed January 22, 2020.
http://hdl.handle.net/11012/40709.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Bílek, Ondřej. “Využití magnetických nanočástic pro transfekci plasmidu do HEK293 buněk
.” 2015. Web. 22 Jan 2020.
Vancouver:
Bílek O. Využití magnetických nanočástic pro transfekci plasmidu do HEK293 buněk
. [Internet] [Thesis]. Brno University of Technology; 2015. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/11012/40709.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Bílek O. Využití magnetických nanočástic pro transfekci plasmidu do HEK293 buněk
. [Thesis]. Brno University of Technology; 2015. Available from: http://hdl.handle.net/11012/40709
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
27.
Oliveira, Jordana Cristina.
Desenvolvimento de estratégias alternativas para teste de fármacos: obtenção e caracterização de linhagens mutantes estáveis de Leishmania expressando luciferase.
Degree: Mestrado, Biologia da Relação Patógeno-Hospedeiro, 2014, University of São Paulo
URL: http://www.teses.usp.br/teses/disponiveis/42/42135/tde-15122014-154829/
;
► A leishmaniose é causada por protozoários do gênero Leishmania e no Brasil, as principais espécies causadoras da leishmaniose cutânea são Leishmania (V.) braziliensis e Leishmania…
(more)
▼ A leishmaniose é causada por protozoários do gênero Leishmania e no Brasil, as principais espécies causadoras da leishmaniose cutânea são Leishmania (V.) braziliensis e Leishmania (L.) amazonenses. O tratamento da leishmaniose apresenta diversas dificuldades, portanto é fundamental a descoberta de novos fármacos ativos, podendo ser detectada em células cultivadas in vitro e também em animais íntegros, através da técnica de bioimageamento. Neste trabalho, propusemo-nos a produzir linhagens de L. (V.) braziliensis e L. (L) amazonenses expressoras de luciferase e caracterizar o comportamento das linhagens mutantes em testes de sensibilidade a fármacos e de infecção in vitro e in vivo. Foi confirmada a emissão de luz pelas linhagens mutantes das duas espécies de Leishmania, em promastigotas e amastigotas. O comportamento das linhagens mutantes obtidas em relação a curvas de crescimento, sensibilidade aos fármacos tamoxifeno e anfoterina B em promastigotas, perfil de infectividade e sobrevivência em macrófagos e sensibilidade de amastigotas à anfotericina B foi comparado ao comportamento das linhagens parentais, não sendo observadas diferenças significativas. Camundongos BALB/c infectados com a linhagem expressora de luciferase de L. (L.) amazonenses desenvolveram lesões comparáveis aos animais infectados com a cepa selvagem, sendo possível quantificar a carga parasitária nesses animais por bioimageamento. Os resultados obtidos neste trabalho indicam que os parasitas mutantes expressores de luciferase obtidos podem ser utilizados em testes de sensibilidade a fármacos tanto in vitro como in vivo, representando um avanço metodológico nessa área de pesquisa.
Leishmaniasis is caused by protozoan parasites in Brazil, the main causative species of cutaneous leishmaniasis are Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonenses. The treatment of leishmaniasis presents several difficulties, and the discovery of new active drugs for the treatment of leishmaniasis is therefore fundamental. The enzyme luciferase is a reporter widely used in screening tests for new drugs. This enzyme catalyzes the oxidation of luciferase in the presence of ATP emitting light that can be detected in cultured cells in vitro as well as in intact animals, using the technique of bioimaging. In this work, we sought to produce strains of L. (V.) braziliensis and L. (L) amazonenses expressing luciferase and characterize the behavior of these mutant strains in drug susceptibility tests and in in vitro and in vivo infections. Production of light was detected in mutants of both species, in all life cycle stages. Mutant strains were compared to their corresponding parental lines as to their growth pattern, infectivity and survival profile in macrophages and sensitivity to amphotericin B and tamoxifen. No significant differences were observed for these parameters. BALB/c mice infected with the luciferase expressing line of L. (L.) amazonenses developed lesions comparable to those in animals infected with the wild-type strain. The parasite…
Advisors/Committee Members: Uliana, Silvia Reni Bortolin.
Subjects/Keywords: Leishmania; Leishmania; Genes repórteres; Luciferase; Luciferase; Reporter genes; Transfecção; Transfection
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APA (6th Edition):
Oliveira, J. C. (2014). Desenvolvimento de estratégias alternativas para teste de fármacos: obtenção e caracterização de linhagens mutantes estáveis de Leishmania expressando luciferase. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/42/42135/tde-15122014-154829/ ;
Chicago Manual of Style (16th Edition):
Oliveira, Jordana Cristina. “Desenvolvimento de estratégias alternativas para teste de fármacos: obtenção e caracterização de linhagens mutantes estáveis de Leishmania expressando luciferase.” 2014. Masters Thesis, University of São Paulo. Accessed January 22, 2020.
http://www.teses.usp.br/teses/disponiveis/42/42135/tde-15122014-154829/ ;.
MLA Handbook (7th Edition):
Oliveira, Jordana Cristina. “Desenvolvimento de estratégias alternativas para teste de fármacos: obtenção e caracterização de linhagens mutantes estáveis de Leishmania expressando luciferase.” 2014. Web. 22 Jan 2020.
Vancouver:
Oliveira JC. Desenvolvimento de estratégias alternativas para teste de fármacos: obtenção e caracterização de linhagens mutantes estáveis de Leishmania expressando luciferase. [Internet] [Masters thesis]. University of São Paulo; 2014. [cited 2020 Jan 22].
Available from: http://www.teses.usp.br/teses/disponiveis/42/42135/tde-15122014-154829/ ;.
Council of Science Editors:
Oliveira JC. Desenvolvimento de estratégias alternativas para teste de fármacos: obtenção e caracterização de linhagens mutantes estáveis de Leishmania expressando luciferase. [Masters Thesis]. University of São Paulo; 2014. Available from: http://www.teses.usp.br/teses/disponiveis/42/42135/tde-15122014-154829/ ;
28.
Aoki, Juliana Ide.
Ação do análogo de purina tóxico tubercidina em Leishmania ssp.
Degree: Mestrado, Fisiopatologia Experimental, 2008, University of São Paulo
URL: http://www.teses.usp.br/teses/disponiveis/5/5160/tde-30092010-130259/
;
► A identificação de genes relacionados com resistência a compostos antiparasitários tem contribuído para um melhor entendimento do mecanismo de ação de alguns desses compostos. Utilizando…
(more)
▼ A identificação de genes relacionados com resistência a compostos antiparasitários tem contribuído para um melhor entendimento do mecanismo de ação de alguns desses compostos. Utilizando a estratégia que permite a indução de super-expressão após transfecção gênica, isolamos dois loci relacionados com resistência ao análogo tóxico de purina, tubercidina (TUB). Em um desses locus identificamos um ortólogo do gene TOR (TOxic nucleoside Resistance) em L. (L.) major (TOR-Lm), capaz de conferir altos níveis de resistência a TUB. A identificação e localização cromossomal do segundo locus foi obtida, mas os testes funcionais em presença de TUB não foram tão significativos quanto os obtidos após a transfecção do TOR-Lm. Na segunda parte desta dissertação avaliamos a eficácia da associação de TUB com um inibidor específico do transporte de nucleosídeos em mamíferos, nitrobenziltioinosina (NBMPR), visando reverter a toxicidade de TUB apenas no hospedeiro. Demonstramos que TUB tem uma potente ação anti-parasitária em culturas de Leishmania spp., e que o inibidor NBMPR é capaz de proteger células mamíferas de camundongos infectados da ação tóxica de TUB.
Gene identification associated with drug resistance has contributed to a better understanding of the mechanism of action of anti parasitic compounds. Using transfection and over-expression selection strategy we isolated two loci related with the resistance of tubercidin (TUB), a toxic analog purine. In the first locus we identified an ortholog of the TOR gene (TOxic nucleoside Resistance) in L. (L.) major (TOR-Lm), capable to render wild cells resistance to TUB after transfection and over-expression. Chromosomal location and identification of the second locus was done, but functional tests in the presence of TUB were not as significant as those obtained after TOR-Lm transfection. In the second part of this work, we evaluate the effectiveness of the association of TUB with an inhibitor specific to the mammals nucleoside transport, as nitrobenzylthioinosine (NBMPR), aimed at reversing the TUB toxicity only on the host. We first demonstrate that TUB has a potent anti-parasitic action in cultures of Leishmania spp. Then, we discuss the capacity of the NBMPR inhibitor to protect infected macrophages from the toxic effects of TUB.
Advisors/Committee Members: Cotrim, Paulo Cesar.
Subjects/Keywords: Leishmania; Leishmania; Nucleosídeos de purina; Purine nucleoside; Transfecção; Transfection; Tubercidin; Tubercidina
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APA (6th Edition):
Aoki, J. I. (2008). Ação do análogo de purina tóxico tubercidina em Leishmania ssp. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/5/5160/tde-30092010-130259/ ;
Chicago Manual of Style (16th Edition):
Aoki, Juliana Ide. “Ação do análogo de purina tóxico tubercidina em Leishmania ssp.” 2008. Masters Thesis, University of São Paulo. Accessed January 22, 2020.
http://www.teses.usp.br/teses/disponiveis/5/5160/tde-30092010-130259/ ;.
MLA Handbook (7th Edition):
Aoki, Juliana Ide. “Ação do análogo de purina tóxico tubercidina em Leishmania ssp.” 2008. Web. 22 Jan 2020.
Vancouver:
Aoki JI. Ação do análogo de purina tóxico tubercidina em Leishmania ssp. [Internet] [Masters thesis]. University of São Paulo; 2008. [cited 2020 Jan 22].
Available from: http://www.teses.usp.br/teses/disponiveis/5/5160/tde-30092010-130259/ ;.
Council of Science Editors:
Aoki JI. Ação do análogo de purina tóxico tubercidina em Leishmania ssp. [Masters Thesis]. University of São Paulo; 2008. Available from: http://www.teses.usp.br/teses/disponiveis/5/5160/tde-30092010-130259/ ;
29.
Aoki, Juliana Ide.
Identificação de um gene que confere resistência a tubercidina em Leishmania (Leishmania) major.
Degree: PhD, Doenças Tropicais e Saúde Internacional, 2013, University of São Paulo
URL: http://www.teses.usp.br/teses/disponiveis/99/99131/tde-21022014-151125/
;
► A identificação de genes relacionados com resistência a compostos antiparasitários tem contribuído para um melhor entendimento do mecanismo de ação de compostos antileishmania. Pouco se…
(more)
▼ A identificação de genes relacionados com resistência a compostos antiparasitários tem contribuído para um melhor entendimento do mecanismo de ação de compostos antileishmania. Pouco se sabe sobre o mecanismo de ação do análogo de purina tubercidina (TUB) em Leishmania. Utilizando a estratégia de superexpressão após transfecção gênica, isolamos um locus de Leishmania (Leishmania) major, de 31 kb, capaz de conferir níveis de resistência quatro vezes maior que o parasita selvagem. Várias deleções desse locus foram geradas e a construção de 3 kb (pSNBR/3kbClaI-EcoRI) também conferiu níveis de resistência quando comparado ao parasita elvagem. Através de análises no genoma de L. (L.) major, localizamos esse locus no cromossomo 31 e, no fragmento de 3 kb, um gene que codifica para uma proteína com função desconhecida até o momento (LmjF.31.2010). Esta proteína foi relacionada com resistência a TUB em todas as linhagens transfectadas analisadas (cosTUB2 e pSNBR/3kbClaI-EcoRI), assim denominamos LmjF.31.2010, de proteína relacionada com resistência a TUB (PRRT). A quantificação relativa de transcritos de mRNA na construção pSNBR/3kbClaI-EcoRI apresentou níveis altos de transcritos da PRRT. Foram gerados ainda mutantes de L. (L.) major e L. (L.) amazonensis resistentes a TUB e estes se apresentaram bem adaptados a concentrações altas de TUB, apresentando razão de resistência maior que 200 vezes, quando comparado com os respectivos parasitas selvagens. A PRRT também foi relacionada na resistência a TUB nos mutantes gerados, pois houve amplificação gênica de prrt. Os resultados obtidos neste trabalho fornecem dados para inferir a importância da PRRT no mecanismo relacionado com resistência a TUB.
The identification of genes associated with resistance to antiparasitic compounds has contributed to a better understanding of the mechanism of action of compounds against Leishmania. Little is known about the mechanism of action of purine analog tubercidin (TUB) in Leishmania. Using a strategy of gene overexpression after transfection, we isolated a locus of Leishmania (Leishmania) major, 31 kb, capable of conferring fold resistance four times greater than the wild type parasite. A set of deletions of this locus were generated and a 3 kb construction (pSNBR/3kbClaI-EcoRI) conferred fold resistance twice than the wild type. Analysis of L. (L.) major genome, located this locus on chromosome 31 and on 3 kb fragment we identified a gene encoding a protein with unknown function (LmjF.31.2010). This protein has been related to TUB resistance in all strains analyzed (cosTUB2 and pSNBR/3kbClaI-EcoRI), so we named mjF.31.2010 of protein related with resistance to TUB (PRRT). Relative quantification of mRNA transcripts in the construction pSNBR/3kbClaI-EcoRI showed high levels of PRRT transcripts.Mutants of L. (L.) major and L. (L.) amazonensis resistant to TUB were also generated and these were well adapted to high TUB concentrations, presenting fol resistance greater than 200 times when compared with their respective wild type. The PRRT…
Advisors/Committee Members: Cotrim, Paulo Cesar.
Subjects/Keywords: Leishmania; Leishmania; Purinas; Purine 4; Transfecção; Transfection; Tubercidin; Tubercidina
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APA ·
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APA (6th Edition):
Aoki, J. I. (2013). Identificação de um gene que confere resistência a tubercidina em Leishmania (Leishmania) major. (Doctoral Dissertation). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/99/99131/tde-21022014-151125/ ;
Chicago Manual of Style (16th Edition):
Aoki, Juliana Ide. “Identificação de um gene que confere resistência a tubercidina em Leishmania (Leishmania) major.” 2013. Doctoral Dissertation, University of São Paulo. Accessed January 22, 2020.
http://www.teses.usp.br/teses/disponiveis/99/99131/tde-21022014-151125/ ;.
MLA Handbook (7th Edition):
Aoki, Juliana Ide. “Identificação de um gene que confere resistência a tubercidina em Leishmania (Leishmania) major.” 2013. Web. 22 Jan 2020.
Vancouver:
Aoki JI. Identificação de um gene que confere resistência a tubercidina em Leishmania (Leishmania) major. [Internet] [Doctoral dissertation]. University of São Paulo; 2013. [cited 2020 Jan 22].
Available from: http://www.teses.usp.br/teses/disponiveis/99/99131/tde-21022014-151125/ ;.
Council of Science Editors:
Aoki JI. Identificação de um gene que confere resistência a tubercidina em Leishmania (Leishmania) major. [Doctoral Dissertation]. University of São Paulo; 2013. Available from: http://www.teses.usp.br/teses/disponiveis/99/99131/tde-21022014-151125/ ;
NSYSU
30.
Lu, Cheng-che.
A Study of Cell-penetrating Peptides as a Delivery Vehicle in Cells of a Model Cnidarian.
Degree: Master, Biological Sciences, 2017, NSYSU
URL: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0711117-105733
► Intracellular symbiosis between Symbiodinium dinoflagellates and corals forms coral reefs providing diverse and essential habitats for the existence of a large variety of marine organisms…
(more)
▼ Intracellular symbiosis between Symbiodinium dinoflagellates and corals forms coral reefs providing diverse and essential habitats for the existence of a large variety of marine organisms possible in the nutrient-poor tropical seas. However, our knowledge on the cellular and molecular mechanisms of this important ecological interaction is poor, partly due to a lack of a genetically manipulatable model of cnidarian. The aim of the present study is to establish a
transfection system in a cnidarian model, Aiptasia pulchella, with cell-penetrating peptides (CPPs) as the delivery vehicle. I have successfully established a CPPs-based protocol to deliver pEGFP-ApHistone reporter plasmid into COS7 cells and expressed EGFP-ApHistone fusion protein in the nucleus. The results showed that the CPPs were able to penetrate A. pulchella cells and replaced the binding between DNA and Hoechst33258, as CPPs were not only penetrated into cells but also diffused into nucleus in the marine cnidarian cell model. Moreover, the results strongly suggests that CPPs carried the Cy3-conjugated DNA primers into the nucleus of A. pulcehlla endodermal cells, revealing its potentials for cell
transfection reagents. Overall, all results provided evidence that the CPPs are one of
transfection candidates for marine cnidarian even the efficiency was low under current protocol. Optimization of proceduces is underway to improve the delivery efficiency.
Advisors/Committee Members: Chang, Hsueh-Wen (committee member), KH Chiu (chair), Lee, Che-Hsin (chair).
Subjects/Keywords: EGFP-ApHistone; cnidarian transfection; ApHistone; cell-penetrating peptides; COS7
Record Details
Similar Records
Cite
Share »
Record Details
Similar Records
Cite
« Share
❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Lu, C. (2017). A Study of Cell-penetrating Peptides as a Delivery Vehicle in Cells of a Model Cnidarian. (Thesis). NSYSU. Retrieved from http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0711117-105733
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Lu, Cheng-che. “A Study of Cell-penetrating Peptides as a Delivery Vehicle in Cells of a Model Cnidarian.” 2017. Thesis, NSYSU. Accessed January 22, 2020.
http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0711117-105733.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Lu, Cheng-che. “A Study of Cell-penetrating Peptides as a Delivery Vehicle in Cells of a Model Cnidarian.” 2017. Web. 22 Jan 2020.
Vancouver:
Lu C. A Study of Cell-penetrating Peptides as a Delivery Vehicle in Cells of a Model Cnidarian. [Internet] [Thesis]. NSYSU; 2017. [cited 2020 Jan 22].
Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0711117-105733.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Lu C. A Study of Cell-penetrating Peptides as a Delivery Vehicle in Cells of a Model Cnidarian. [Thesis]. NSYSU; 2017. Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0711117-105733
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
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Open Access Theses and Dissertations