subject:(gene targeting)

University of Texas Southwestern Medical Center

1. Ellis, Brian Lee. Improving Viral Vectors for Gene Targeting in Gene Therapy.

Degree: 2011, University of Texas Southwestern Medical Center

► Over 10,000 monogenic diseases in the world affect one out of every hundred live births (WHO). Gene targeting is a term that is used describe… (more)

Subjects/Keywords: Gene Targeting; Gene Therapy; Lentivirus

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APA (6^th Edition):

Ellis, B. L. (2011). Improving Viral Vectors for Gene Targeting in Gene Therapy. (Thesis). University of Texas Southwestern Medical Center. Retrieved from http://hdl.handle.net/2152.5/837

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Ellis, Brian Lee. “Improving Viral Vectors for Gene Targeting in Gene Therapy.” 2011. Thesis, University of Texas Southwestern Medical Center. Accessed November 18, 2019. http://hdl.handle.net/2152.5/837.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Ellis, Brian Lee. “Improving Viral Vectors for Gene Targeting in Gene Therapy.” 2011. Web. 18 Nov 2019.

Vancouver:

Ellis BL. Improving Viral Vectors for Gene Targeting in Gene Therapy. [Internet] [Thesis]. University of Texas Southwestern Medical Center; 2011. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/2152.5/837.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Ellis BL. Improving Viral Vectors for Gene Targeting in Gene Therapy. [Thesis]. University of Texas Southwestern Medical Center; 2011. Available from: http://hdl.handle.net/2152.5/837

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Utah

2. Morton, John Jason. Zinc finger nuclease-induced double-strand breaks mediate targeted mutagenesis in Caenorhabditis elegans;.

Degree: PhD, Biochemistry;, 2007, University of Utah

URL: http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1121/rec/1477

► Zinc finger nucleases (ZFNs) are chimeric proteins composed of a DNA-binding domain comprised of several tandem Cys2His2 zinc fingers and a nonspecific endonuclease domain from… (more)

Subjects/Keywords: Chimeric Proteins; Gene Targeting

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APA (6^th Edition):

Morton, J. J. (2007). Zinc finger nuclease-induced double-strand breaks mediate targeted mutagenesis in Caenorhabditis elegans;. (Doctoral Dissertation). University of Utah. Retrieved from http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1121/rec/1477

Chicago Manual of Style (16^th Edition):

Morton, John Jason. “Zinc finger nuclease-induced double-strand breaks mediate targeted mutagenesis in Caenorhabditis elegans;.” 2007. Doctoral Dissertation, University of Utah. Accessed November 18, 2019. http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1121/rec/1477.

MLA Handbook (7^th Edition):

Morton, John Jason. “Zinc finger nuclease-induced double-strand breaks mediate targeted mutagenesis in Caenorhabditis elegans;.” 2007. Web. 18 Nov 2019.

Vancouver:

Morton JJ. Zinc finger nuclease-induced double-strand breaks mediate targeted mutagenesis in Caenorhabditis elegans;. [Internet] [Doctoral dissertation]. University of Utah; 2007. [cited 2019 Nov 18]. Available from: http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1121/rec/1477.

Council of Science Editors:

Morton JJ. Zinc finger nuclease-induced double-strand breaks mediate targeted mutagenesis in Caenorhabditis elegans;. [Doctoral Dissertation]. University of Utah; 2007. Available from: http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1121/rec/1477

University of Hong Kong

3. Chan, Fu-lun. Effective DNA delivery mediated by pH responsive peptides.

Degree: Master of Medical Sciences, 2012, University of Hong Kong

URL: Chan, F. [陳賦麟]. (2012). Effective DNA delivery mediated by pH responsive peptides. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4833333 ; http://dx.doi.org/10.5353/th_b4833333 ; http://hdl.handle.net/10722/173921

►

Non-viral vectors have been used to deliver therapeutic genes to treat different diseases. There are a variety of non-viral vectors such as liposomes, cationic polymers… (more)

Subjects/Keywords: Peptides.; Gene targeting.; Genetic vectors.; Gene therapy.

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APA (6^th Edition):

Chan, F. (2012). Effective DNA delivery mediated by pH responsive peptides. (Masters Thesis). University of Hong Kong. Retrieved from Chan, F. [陳賦麟]. (2012). Effective DNA delivery mediated by pH responsive peptides. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4833333 ; http://dx.doi.org/10.5353/th_b4833333 ; http://hdl.handle.net/10722/173921

Chicago Manual of Style (16^th Edition):

Chan, Fu-lun. “Effective DNA delivery mediated by pH responsive peptides.” 2012. Masters Thesis, University of Hong Kong. Accessed November 18, 2019. Chan, F. [陳賦麟]. (2012). Effective DNA delivery mediated by pH responsive peptides. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4833333 ; http://dx.doi.org/10.5353/th_b4833333 ; http://hdl.handle.net/10722/173921.

MLA Handbook (7^th Edition):

Chan, Fu-lun. “Effective DNA delivery mediated by pH responsive peptides.” 2012. Web. 18 Nov 2019.

Vancouver:

Chan F. Effective DNA delivery mediated by pH responsive peptides. [Internet] [Masters thesis]. University of Hong Kong; 2012. [cited 2019 Nov 18]. Available from: Chan, F. [陳賦麟]. (2012). Effective DNA delivery mediated by pH responsive peptides. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4833333 ; http://dx.doi.org/10.5353/th_b4833333 ; http://hdl.handle.net/10722/173921.

Council of Science Editors:

Chan F. Effective DNA delivery mediated by pH responsive peptides. [Masters Thesis]. University of Hong Kong; 2012. Available from: Chan, F. [陳賦麟]. (2012). Effective DNA delivery mediated by pH responsive peptides. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4833333 ; http://dx.doi.org/10.5353/th_b4833333 ; http://hdl.handle.net/10722/173921

University of Texas Southwestern Medical Center

4. Checketts, Joshua Allen. Nuclease-Mediated Targeted Gene Insertion at the Adenosine Deaminase Locus in Primary Cells.

Degree: 2013, University of Texas Southwestern Medical Center

URL: http://hdl.handle.net/2152.5/1727

► Gene therapy is the ability to correct diseases at the DNA level and has long been a goal of science and medicine. The earliest gene… (more)

▼ Gene therapy is the ability to correct diseases at the DNA level and has long been a goal of science and medicine. The earliest gene therapy clinical trial was for a patient with severe combined immunodeficiency (SCID) due to adenosine deaminase (ADA) deficiency. Initial trials looked promising and the technique was extended to other forms of primary immunodeficiency. Unfortunately, some of the patients enrolled in these trials using retroviral vectors to carry replacement genes resulted in insertional oncogenesis. To avoid the insertional oncogenesis caused by random integration into the genome, we postulated that targeted insertion of the gene of interest through homologous recombination would prove to be a safer alternative to random viral insertion of a gene. To this end, we developed several pairs of TAL effector nucleases (TALENs) designed to target exon 1 of ADA. These TALENs function as dimers, and each pair creates a different targeted double strand break near the start site of the ADA gene. The most effective pair induces a DNA double strand break immediately preceding the ADA start codon. Targeted activity of these TALENs was measured through determining the percent of alleles that undergo mutagenic non-homologous end joining upon exposure to the TALENs, with up to 14% of alleles undergoing such mutations. In order to stimulate gene targeting at the ADA locus in human cells, these TALENs were nucleofected into the cells as plasmid DNA, along with a donor plasmid that contains the DNA to be inserted flanked by 800bp arms of homology to the cut site. These TALENs were able to stimulate site-specific integration of the desired fragment at rates of up to 10% in human cell lines. Successful targeted gene insertion was verified through maintained fluorescence, western blots, and sequencing of the targeted alleles through PCR amplification. We demonstrated the ability to enrich for targeted cells through the expression of a selectable marker within the DNA cassette integrated at the ADA locus. In addition to the editing of cell lines, we showed successful stimulation of gene targeting in patient-derived fibroblasts in 1.5% of cells. We demonstrated the feasibility of using the ADA locus as a safe harbor through the targeted insertion of three therapeutically interesting genes. Finally, we demonstrated the successful targeted gene insertion in human CD34+ in up to 0.5% of cells treated. The successful targeting of human CD34+ is especially relevant, as these cells will need to undergo gene targeting in order to be therapeutically relevant as a curative therapy for SCID due to ADA deficiency. Advisors/Committee Members: Albanesi, Joseph P., Porteus, Matthew H., Burma, Sandeep, Abrams, John M., Sternweis, Paul C..

Subjects/Keywords: Gene Therapy; Gene Targeting; Severe Combined Immunodeficiency

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APA (6^th Edition):

Checketts, J. A. (2013). Nuclease-Mediated Targeted Gene Insertion at the Adenosine Deaminase Locus in Primary Cells. (Thesis). University of Texas Southwestern Medical Center. Retrieved from http://hdl.handle.net/2152.5/1727

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Checketts, Joshua Allen. “Nuclease-Mediated Targeted Gene Insertion at the Adenosine Deaminase Locus in Primary Cells.” 2013. Thesis, University of Texas Southwestern Medical Center. Accessed November 18, 2019. http://hdl.handle.net/2152.5/1727.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Checketts, Joshua Allen. “Nuclease-Mediated Targeted Gene Insertion at the Adenosine Deaminase Locus in Primary Cells.” 2013. Web. 18 Nov 2019.

Vancouver:

Checketts JA. Nuclease-Mediated Targeted Gene Insertion at the Adenosine Deaminase Locus in Primary Cells. [Internet] [Thesis]. University of Texas Southwestern Medical Center; 2013. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/2152.5/1727.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Checketts JA. Nuclease-Mediated Targeted Gene Insertion at the Adenosine Deaminase Locus in Primary Cells. [Thesis]. University of Texas Southwestern Medical Center; 2013. Available from: http://hdl.handle.net/2152.5/1727

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Georgia Tech

5. Ruff, Patrick. Protein-assisted targeting of genes in yeast and human cells.

Degree: PhD, Biology, 2013, Georgia Tech

URL: http://hdl.handle.net/1853/52907

► This work was designed as a proof-of-principle concept or prototype to show the effect of protein-assisted targeting of DNA to specific genomic loci. Two strategies… (more)

▼ This work was designed as a proof-of-principle concept or prototype to show the effect of protein-assisted targeting of DNA to specific genomic loci. Two strategies were employed to deliver the DNA with the aim that once inside the cell the DNA would be delivered to the target sequence by the assistance of a protein. In our case, the chosen protein was the site-specific meganuclease I-SceI. The first strategy described herein was to bind the targeting DNA to I-SceI by the use of a fusion protein between I-SceI and a known DNA-binding domain, the GAL4-DBD. The second strategy involved using a DNA aptamer to I-SceI to link the targeting DNA and I-SceI. Testing in vivo revealed that in our human cells (HEK-293) single-stranded DNA was more efficient at gene targeting than double-stranded DNA. In order for the first strategy to work, we needed to have some region of double-stranded DNA. We found that in human cells, it was better for gene targeting to have that double-stranded DNA on the 5’ side of our targeting DNA. We also used gel shift assays to confirm binding by our candidate DNA-binding domain, the GAL4-DBD. We were unable to detect expression of the fusion protein of I-SceI and the GAL4-DBD. For the second strategy we were able to construct an aptamer to I-SceI using a variant of the systematic evolution of ligands by exponential enrichment (SELEX). The I-SceI aptamer was synthesized as part of a longer DNA molecule containing homology to a target locus. Using this chimeric oligonucleotide (part aptamer, part DNA repair region) testing was done in both yeast and human cells. Aside from instances where the aptamer’s secondary structure may have been compromised, the aptamer containing oligonucleotide stimulated repair at a rate 2 to 15-fold higher than the non-selected control sequence. These experimental results show that by delivering targeting DNA within close proximity to the site of modification, gene targeting frequencies can be increased. Advisors/Committee Members: Storici, Francesca (advisor), Chernoff, Yury (committee member), Lieberman, Raquel (committee member), Lobachev, Kirill (committee member), Fan, Yuhong (committee member).

Subjects/Keywords: Aptamer; SELEX; I-SceI; Gene targeting; Protein-assisted targeting

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APA (6^th Edition):

Ruff, P. (2013). Protein-assisted targeting of genes in yeast and human cells. (Doctoral Dissertation). Georgia Tech. Retrieved from http://hdl.handle.net/1853/52907

Chicago Manual of Style (16^th Edition):

Ruff, Patrick. “Protein-assisted targeting of genes in yeast and human cells.” 2013. Doctoral Dissertation, Georgia Tech. Accessed November 18, 2019. http://hdl.handle.net/1853/52907.

MLA Handbook (7^th Edition):

Ruff, Patrick. “Protein-assisted targeting of genes in yeast and human cells.” 2013. Web. 18 Nov 2019.

Vancouver:

Ruff P. Protein-assisted targeting of genes in yeast and human cells. [Internet] [Doctoral dissertation]. Georgia Tech; 2013. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/1853/52907.

Council of Science Editors:

Ruff P. Protein-assisted targeting of genes in yeast and human cells. [Doctoral Dissertation]. Georgia Tech; 2013. Available from: http://hdl.handle.net/1853/52907

The Ohio State University

6. Stachler, Matthew D. Design and engineering of capsid modified AAV-Based vectors targeted towards angiogenic and proliferating vasculature.

Degree: PhD, Integrated Biomedical Science, 2007, The Ohio State University

URL: http://rave.ohiolink.edu/etdc/view?acc_num=osu1180370373

► Angiogenesis and vascular proliferation are involved in a wide variety of diseases ranging from cardiovascular ischemia to cancer. The ability to deliver a therapeutic gene… (more)

▼ Angiogenesis and vascular proliferation are involved in a wide variety of diseases ranging from cardiovascular ischemia to cancer. The ability to deliver a therapeutic gene to these proliferating endothelial cells (EC) could be of great therapeutic value. Adeno-associated virus (AAV) vectors transduce a wide variety of cell types; however, cells of the vascular system remain refractive to AAV-mediated gene transduction. Capsids of different serotypes have been used to try to increase the ability of AAV vectors to transduce the vascular endothelium. While still limiting, AAV serotype 1-based vectors have shown the most promise. Another method to increase transduction on cells not normally permissible to easy transduction is to incorporate small targeting peptides into the vector capsid. To improve upon AAV1’s vascular tropism, peptides targeted to receptors expressed on the vasculature were incorporated into the AAV1 capsid. Modified vectors were used to transduce four different primary human EC populations. Several modified vectors, including vectors targeted to vascular endothelial growth factor receptor 2 (VEGFR2), Tie2, and alphav beta3/5 integrins, significantly increased transduction of all ECs tested (average 10 to 60-fold). Cellular binding, internalization, and intracellular trafficking was further characterized in these vectors. It was found that transduction of vascular endothelial cells was increased by different mechanisms in the vectors targeting the different receptors. The Tie2 targeted vector increased the amount of vector bound to the cell surface, but required internalization through AAV1’s natural mechanism. The VEGFR2 targeted vector required initial binding to AAV1’s primary receptor sialic acid, but intracellular trafficking differed from untargeted AAV1. Transduction of the ECs was completely independent of sialic acid using the integrin targeted vector. To test the ability of these vectors to target angiogenic vasculature in vivo, a murine xenograph model of human ovarian cancer was utilized. A human ovarian cancer cell line was injected intraperitoneal, allowed to grow and promote angiogenesis, and vector delivering a green fluorescent protein-luciferase fusion transgene was injected intravenously through the tail vein. The integrin targeted and one of the VEGFR2 targeted vectors were found to significantly increase gene transfer to the tumor enviroment by live animal bioluminescence. Advisors/Committee Members: Bartlett, Jeffrey (Advisor).

Subjects/Keywords: Gene Therapy; Adeno-Associated Virus; Targeting; Angiogenesis

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APA (6^th Edition):

Stachler, M. D. (2007). Design and engineering of capsid modified AAV-Based vectors targeted towards angiogenic and proliferating vasculature. (Doctoral Dissertation). The Ohio State University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=osu1180370373

Chicago Manual of Style (16^th Edition):

Stachler, Matthew D. “Design and engineering of capsid modified AAV-Based vectors targeted towards angiogenic and proliferating vasculature.” 2007. Doctoral Dissertation, The Ohio State University. Accessed November 18, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1180370373.

MLA Handbook (7^th Edition):

Stachler, Matthew D. “Design and engineering of capsid modified AAV-Based vectors targeted towards angiogenic and proliferating vasculature.” 2007. Web. 18 Nov 2019.

Vancouver:

Stachler MD. Design and engineering of capsid modified AAV-Based vectors targeted towards angiogenic and proliferating vasculature. [Internet] [Doctoral dissertation]. The Ohio State University; 2007. [cited 2019 Nov 18]. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1180370373.

Council of Science Editors:

Stachler MD. Design and engineering of capsid modified AAV-Based vectors targeted towards angiogenic and proliferating vasculature. [Doctoral Dissertation]. The Ohio State University; 2007. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=osu1180370373

University of Hong Kong

7. Cao, Shanbo. Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis.

Degree: PhD, 2012, University of Hong Kong

URL: Cao, S. [曹善柏]. (2012). Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4979925 ; http://dx.doi.org/10.5353/th_b4979925 ; http://hdl.handle.net/10722/207896

► Spermatogenesis is regulated by steroid hormones which induce expression of various genes responsible for the growth, proliferation and differentiation of spermatogonia to form mature haploid… (more)

▼ Spermatogenesis is regulated by steroid hormones which induce expression of various genes responsible for the growth, proliferation and differentiation of spermatogonia to form mature haploid spermatozoa. The surrounding somatic cells including Leydig and Sertoli cells support the whole process in vivo. Previously, we used the post-vitamin A treated vitamin-A-deficient (PVA-VAD) rat model to study spermatogenesis, and identified 24 genes that are differentially up-regulated after retinol treatment. Vad1.2 is one of the up-regulated transcripts expressed in the rat testis from postnatal day 25. Vad1.2 transcript is localized to the round and elongating spermatids in the adult mouse testis. In silico analyses showed that Vad1.2 transcript is down-regulated in patients with teratozoospermia and non-obstructive azoospermia, suggesting that Vad1.2 may have important roles in spermatogenesis. However, how Vad1.2 affects spermatogenesis remains unclear. Therefore, the present study was designed to study the functional roles of Vad1.2 protein in mice using gene targeting approach and investigate the molecular changes in mice with Vad1.2 deficiency. Vad1.2 polyclonal antibody was raised against the full-length mouse Vad1.2 recombination protein and affinity purified. Vad1.2 protein was localized to the cytoplasm and flagellum of condensing spermatids, specifically to the fibrous sheath (FS) in cauda epididymal spermatozoa. Vad1.2 conditional knockout vector was constructed and used to generate Vad1.2 null mice. Vad1.2-/- male mice developed normally but were subfertile with reduced sperm count and motility. Vad1.2-/- male mice had smaller testis and higher incident of sloughing of immature germ cells into the seminiferous lumens when compared to the wild-type. Yet, the rates of germ cell proliferation and apoptosis were similar between the wild-type and the mutant testis. Interestingly approximately 50% of the mutant cauda epididymal spermatozoa showed deformed flagella and demonstrated structural defects typically associated with bending of flagellum at the principal piece or at the midpiece/principal piece junctions. The acrosome, nucleus and mitochondrial sheath of these spermatozoa appeared normal, while the flagellum displayed structural abnormalities including deformation of the two longitudinal columns of the FS and disruption of a portion of FS, suggesting that Vad1.2 might be involved in the biogenesis of FS in spermatogenesis. Furthermore, Vad1.2 interacted with Akap4 in vivo, and the two proteins were co-immunoprecipitated from the testis or cauda epididymal spermatozoa lysates. Akap4 and Vad1.2 were localized to the tail region of the testicular spermatids and cauda epididymal spermatozoa. The expression levels of pro- and mature Akap4 in Vad1.2-/- testes were markedly increased when compared with the wild-type mice. However, a significant decrease of Akap4 was found in the mutant cauda epididymal spermatozoa, suggesting that most of the mature Akap4 failed to incorporate into the FS. Taken together, Vad1.2 plays… Advisors/Committee Members: Yeung, WSB, Lee, CKF.

Subjects/Keywords: Gene targeting; Spermatogenesis in animals - Genetic aspects

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APA (6^th Edition):

Cao, S. (2012). Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. (Doctoral Dissertation). University of Hong Kong. Retrieved from Cao, S. [曹善柏]. (2012). Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4979925 ; http://dx.doi.org/10.5353/th_b4979925 ; http://hdl.handle.net/10722/207896

Chicago Manual of Style (16^th Edition):

Cao, Shanbo. “Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis.” 2012. Doctoral Dissertation, University of Hong Kong. Accessed November 18, 2019. Cao, S. [曹善柏]. (2012). Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4979925 ; http://dx.doi.org/10.5353/th_b4979925 ; http://hdl.handle.net/10722/207896.

MLA Handbook (7^th Edition):

Cao, Shanbo. “Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis.” 2012. Web. 18 Nov 2019.

Vancouver:

Cao S. Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. [Internet] [Doctoral dissertation]. University of Hong Kong; 2012. [cited 2019 Nov 18]. Available from: Cao, S. [曹善柏]. (2012). Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4979925 ; http://dx.doi.org/10.5353/th_b4979925 ; http://hdl.handle.net/10722/207896.

Council of Science Editors:

Cao S. Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. [Doctoral Dissertation]. University of Hong Kong; 2012. Available from: Cao, S. [曹善柏]. (2012). Gene targeting to study a novel testis-specific gene Vad1.2 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4979925 ; http://dx.doi.org/10.5353/th_b4979925 ; http://hdl.handle.net/10722/207896

University of Hong Kong

8. Zuo, Yan. Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis.

Degree: PhD, 2008, University of Hong Kong

URL: Zuo, Y. [左妍]. (2008). Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4163425 ; http://dx.doi.org/10.5353/th_b4163425 ; http://hdl.handle.net/10722/54422

published_or_final_version

Obstetrics and Gynaecology

Doctoral

Doctor of Philosophy

Advisors/Committee Members: Lee, CKF, Yeung, WSB.

Subjects/Keywords: Spermatogenesis.; Gene targeting.

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APA (6^th Edition):

Zuo, Y. (2008). Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. (Doctoral Dissertation). University of Hong Kong. Retrieved from Zuo, Y. [左妍]. (2008). Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4163425 ; http://dx.doi.org/10.5353/th_b4163425 ; http://hdl.handle.net/10722/54422

Chicago Manual of Style (16^th Edition):

Zuo, Yan. “Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis.” 2008. Doctoral Dissertation, University of Hong Kong. Accessed November 18, 2019. Zuo, Y. [左妍]. (2008). Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4163425 ; http://dx.doi.org/10.5353/th_b4163425 ; http://hdl.handle.net/10722/54422.

MLA Handbook (7^th Edition):

Zuo, Yan. “Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis.” 2008. Web. 18 Nov 2019.

Vancouver:

Zuo Y. Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. [Internet] [Doctoral dissertation]. University of Hong Kong; 2008. [cited 2019 Nov 18]. Available from: Zuo, Y. [左妍]. (2008). Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4163425 ; http://dx.doi.org/10.5353/th_b4163425 ; http://hdl.handle.net/10722/54422.

Council of Science Editors:

Zuo Y. Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. [Doctoral Dissertation]. University of Hong Kong; 2008. Available from: Zuo, Y. [左妍]. (2008). Gene targeting to study a novel acrosome specific gene VAD1.3/AEP1 in spermatogenesis. (Thesis). University of Hong Kong, Pokfulam, Hong Kong SAR. Retrieved from http://dx.doi.org/10.5353/th_b4163425 ; http://dx.doi.org/10.5353/th_b4163425 ; http://hdl.handle.net/10722/54422

University of Southern California

9. Yaegashi, Junko. Application of genome-wide strategies for the mining of secondary metabolite biosynthesis pathways in filamentous fungi.

Degree: PhD, Pharmaceutical Sciences, 2017, University of Southern California

URL: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/553827/rec/854

► Genome projects of filamentous fungi have generated an unprecedented amount of information, and in a moment in time often referred to as the “post-genomic era”,… (more)

▼ Genome projects of filamentous fungi have generated an unprecedented amount of information, and in a moment in time often referred to as the “post-genomic era”, it is up to us to find ways to provide meaning to this immense quantity of genome sequence data. Fungi have long been known to be prolific producers of bioactive secondary metabolites, but bioinformatic analysis has showed that these organisms harbor the potential to produce far more secondary metabolites than are currently known. The work herein describes genome-wide strategies to approach two main challenges we now face: 1) finding ways to access the hidden natural products and 2) developing and/ or using genetic systems in fungal species to link secondary metabolites to their biosynthesis genes and elucidate their biosynthesis pathways. We have demonstrated the power of combining bioinformatics, molecular gene targeting, and natural product chemistry in secondary metabolite biosynthesis research. ❧ We initally used A. nidulans as the target species and took advantage of its well-established gene targeting system. First, genome-based deletion analysis in A. nidulans led us to find genes involved in the biosynthesis of xanthones located in at least three distinct loci in the genome. This was particularly interesting because it contradicted the general notion that fungal secondary metabolites are clustered. This highlighted the utility of genomics combined with efficient gene targeting to identify these dispersed genes. Next, we hypothesized that altering the expression of kinases would have an effect on secondary metabolite production and may activate silent gene clusters. We used a genome-wide kinase knockout library and screened its secondary metabolite profile and found that the mpkA deletion strain produced aspernidine A, a compound not previously isolated from A. nidulans. We performed additional gene deletions and determined the border of the biosynthesis gene cluster and proposed the biosynthesis pathway for aspernidine A. ❧ We then took advantage of the recent advances in genome sequencing of Penicillium species and chose Penicillium canescens as the target species. This species is a prolific producer of secondary metabolites but no previous work has been reported to link their metabolites to biosynthesis genes. This is due in large part to the lack of an efficient gene targeting system in this organism. We therefore developed a genetic system in P. canescens that would allow us to perform targeted gene manipulations rapidly and efficiently. We then applied this system and generated a genome-wide deletion library of polyketide synthase (PKS) genes and nonribosomal peptide synthetase (NRPS) genes. Using this library, we successfully linked four secondary metabolites, griseofulvin, xanthoepocin, 15-deoxyoxalicine B, and amauromine, to their core secondary metabolite biosynthesis genes. Interestingly, 15-deoxyoxalicine B is a structurally unique diterpenic meroterpenoid, a class of fungal metabolites whose biosynthesis mechanism remains largely unknown.… Advisors/Committee Members: Wang, Clay C.C. (Committee Chair), Stiles, Bangyan L. (Committee Member), Olenyuk, Bogdan (Committee Member), Okamoto, Curtis Toshio (Committee Member), Zhang, Chao (Committee Member).

Subjects/Keywords: Aspergillus; Penicillium; genome mining; gene targeting

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Yaegashi, J. (2017). Application of genome-wide strategies for the mining of secondary metabolite biosynthesis pathways in filamentous fungi. (Doctoral Dissertation). University of Southern California. Retrieved from http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/553827/rec/854

Chicago Manual of Style (16^th Edition):

Yaegashi, Junko. “Application of genome-wide strategies for the mining of secondary metabolite biosynthesis pathways in filamentous fungi.” 2017. Doctoral Dissertation, University of Southern California. Accessed November 18, 2019. http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/553827/rec/854.

MLA Handbook (7^th Edition):

Yaegashi, Junko. “Application of genome-wide strategies for the mining of secondary metabolite biosynthesis pathways in filamentous fungi.” 2017. Web. 18 Nov 2019.

Vancouver:

Yaegashi J. Application of genome-wide strategies for the mining of secondary metabolite biosynthesis pathways in filamentous fungi. [Internet] [Doctoral dissertation]. University of Southern California; 2017. [cited 2019 Nov 18]. Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/553827/rec/854.

Council of Science Editors:

Yaegashi J. Application of genome-wide strategies for the mining of secondary metabolite biosynthesis pathways in filamentous fungi. [Doctoral Dissertation]. University of Southern California; 2017. Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll3/id/553827/rec/854

Hong Kong University of Science and Technology

10. Guo, Weilin LIFS. Utilization of non-homologous end joining (NHEJ) mediated knock-in method via CRISPR/Cas9 system to achieve visible conditional knock-out in zebrafish.

Degree: 2016, Hong Kong University of Science and Technology

URL: https://doi.org/10.14711/thesis-b1610645 ; http://repository.ust.hk/ir/bitstream/1783.1-95287/1/th_redirect.html

► In biological studies, insertion of exogenous sequences, such as protein tags, fluorescent reporters or loxp sites into targeted genomic locus provides a powerful tool to… (more)

Subjects/Keywords: Genetic engineering; Gene targeting; Zebra danio; Genetics

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APA (6^th Edition):

Guo, W. L. (2016). Utilization of non-homologous end joining (NHEJ) mediated knock-in method via CRISPR/Cas9 system to achieve visible conditional knock-out in zebrafish. (Thesis). Hong Kong University of Science and Technology. Retrieved from https://doi.org/10.14711/thesis-b1610645 ; http://repository.ust.hk/ir/bitstream/1783.1-95287/1/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 (16^th Edition):

Guo, Weilin LIFS. “Utilization of non-homologous end joining (NHEJ) mediated knock-in method via CRISPR/Cas9 system to achieve visible conditional knock-out in zebrafish.” 2016. Thesis, Hong Kong University of Science and Technology. Accessed November 18, 2019. https://doi.org/10.14711/thesis-b1610645 ; http://repository.ust.hk/ir/bitstream/1783.1-95287/1/th_redirect.html.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Guo, Weilin LIFS. “Utilization of non-homologous end joining (NHEJ) mediated knock-in method via CRISPR/Cas9 system to achieve visible conditional knock-out in zebrafish.” 2016. Web. 18 Nov 2019.

Vancouver:

Guo WL. Utilization of non-homologous end joining (NHEJ) mediated knock-in method via CRISPR/Cas9 system to achieve visible conditional knock-out in zebrafish. [Internet] [Thesis]. Hong Kong University of Science and Technology; 2016. [cited 2019 Nov 18]. Available from: https://doi.org/10.14711/thesis-b1610645 ; http://repository.ust.hk/ir/bitstream/1783.1-95287/1/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:

Guo WL. Utilization of non-homologous end joining (NHEJ) mediated knock-in method via CRISPR/Cas9 system to achieve visible conditional knock-out in zebrafish. [Thesis]. Hong Kong University of Science and Technology; 2016. Available from: https://doi.org/10.14711/thesis-b1610645 ; http://repository.ust.hk/ir/bitstream/1783.1-95287/1/th_redirect.html

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Notre Dame

11. Zhi Xu. Dissection of the Multiple Functions of Plasminogen Activator Inhibitor-1</h1>.

Degree: PhD, Chemistry and Biochemistry, 2008, University of Notre Dame

URL: https://curate.nd.edu/show/9w032229q44

► Plasminogen Activator Inhibitor-1 (PAI-1) is the main physiological regulator of tissue-type plasminogen activator (tPA) in normal plasma. In addition to its critical function in… (more)

▼ Plasminogen Activator Inhibitor-1 (PAI-1) is the main physiological regulator of tissue-type plasminogen activator (tPA) in normal plasma. In addition to its critical function in fibrinolysis, PAI-1 has been implicated in other physiological and pathophysiological processes. Interestingly, both antifibrinolytic-related and non-antifibrinolytic-related roles of PAI-1 function have been implicated in the process of angiogenesis, which might account for some discrepancies observed in angiogenic models using PAI-1 deficient and over-expressing transgenic mice. To investigate the structure-function relationships of mouse PAI-1, the recombinant PAI-1 proteins were expressed in Escherichia coli and characterized. Our studies indicated that the complex interactions traditionally associated with different functions of human PAI-1 apply to the murine system, thus demonstrating a commonality of subtle functions among different species and evolutionary conservation of this protein. In an effort to separate these functions in vivo, knock-in targeting vector with targeted mutations to ablate the vitronectin binding ability was constructed. The composition of the targeting vector includes the 5’ and 3’ flanking sequences containing the mutation, a LoxP-FLT-Neo-FLT-LoxP cassette, and a CDA expression cassette. After homologous recombination with the WT allele, the “floxed” cassette may be removed by crossing with either Cre recombinase or flipase expressing transgenic mice. A cell culture-free system was developed to examine the cre-recombinase-mediated excision of the “floxed” element before the targeting vector was introduced into the embryonic stem cells. In order to bypass the lengthy backcrossing process from the 129 background into the C57BL/6 background, the homologous arms in the targeting vector were constructed using a C57BL/6 genetic background to facilitate the backcrossing. The targeting vectors were introduced into both C57 and C57/129 hybrid embryonic stem cells by electroporation, and the homologous recombination was screened by Southern Blot analysis. Positive ES cell clones with homologous recombination at both arms were identified. These PAI-1 knock-in mice would serve to further elucidate mechanisms associated with the observed phenotypes in PAI-1 deficient mice in a number of challenge models. Advisors/Committee Members: Dr. Francis J. Castellino, Committee Member, Dr. Holly Goodson, Committee Member, Dr. Jeffrey S.Schorey, Committee Member, Dr. Dinshaw S Balsara , Committee Chair, Dr. Anthony S. Serianni, Committee Member.

Subjects/Keywords: coagulation; gene targeting

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Xu, Z. (2008). Dissection of the Multiple Functions of Plasminogen Activator Inhibitor-1</h1>. (Doctoral Dissertation). University of Notre Dame. Retrieved from https://curate.nd.edu/show/9w032229q44

Chicago Manual of Style (16^th Edition):

Xu, Zhi. “Dissection of the Multiple Functions of Plasminogen Activator Inhibitor-1</h1>.” 2008. Doctoral Dissertation, University of Notre Dame. Accessed November 18, 2019. https://curate.nd.edu/show/9w032229q44.

MLA Handbook (7^th Edition):

Xu, Zhi. “Dissection of the Multiple Functions of Plasminogen Activator Inhibitor-1</h1>.” 2008. Web. 18 Nov 2019.

Vancouver:

Xu Z. Dissection of the Multiple Functions of Plasminogen Activator Inhibitor-1</h1>. [Internet] [Doctoral dissertation]. University of Notre Dame; 2008. [cited 2019 Nov 18]. Available from: https://curate.nd.edu/show/9w032229q44.

Council of Science Editors:

Xu Z. Dissection of the Multiple Functions of Plasminogen Activator Inhibitor-1</h1>. [Doctoral Dissertation]. University of Notre Dame; 2008. Available from: https://curate.nd.edu/show/9w032229q44

University of Toronto

12. Nelson, Seana ML. Generating C-peptide Humanized Mice.

Degree: 2012, University of Toronto

URL: http://hdl.handle.net/1807/69989

►

Type 1 diabetes is primarily caused by the loss of insulin producing beta-cells. Future therapies based on transplantation of in vitro generated beta-cells hold great… (more)

Subjects/Keywords: Mouse model; Gene targeting; 0369; 0307

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APA (6^th Edition):

Nelson, S. M. (2012). Generating C-peptide Humanized Mice. (Masters Thesis). University of Toronto. Retrieved from http://hdl.handle.net/1807/69989

Chicago Manual of Style (16^th Edition):

Nelson, Seana ML. “Generating C-peptide Humanized Mice.” 2012. Masters Thesis, University of Toronto. Accessed November 18, 2019. http://hdl.handle.net/1807/69989.

MLA Handbook (7^th Edition):

Nelson, Seana ML. “Generating C-peptide Humanized Mice.” 2012. Web. 18 Nov 2019.

Vancouver:

Nelson SM. Generating C-peptide Humanized Mice. [Internet] [Masters thesis]. University of Toronto; 2012. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/1807/69989.

Council of Science Editors:

Nelson SM. Generating C-peptide Humanized Mice. [Masters Thesis]. University of Toronto; 2012. Available from: http://hdl.handle.net/1807/69989

13. Pinho, Raquel de Mello e. Uso do silenciamento gênico mediado por RNA de interferência e de TAL effector nucleases para aumento de eventos gene targeting em células de cão.

Degree: Mestrado, Anatomia dos Animais Domésticos e Silvestres, 2014, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/10/10132/tde-13012015-102643/ ;

►

A inserção de DNA exógeno no genoma hospedeiro é conseguida principalmente através da utilização de vias de reparo como a junção de pontas não homólogas,… (more)

▼

A inserção de DNA exógeno no genoma hospedeiro é conseguida principalmente através da utilização de vias de reparo como a junção de pontas não homólogas, que possui caráter aleatório, e a recombinação homóloga, que possibilita o gene targeting. Algumas ferramentas como as TAL Effector Nucleases (TALENs) e o RNA interferência (RNAi) podem ser utilizadas para aumentar a taxa de integração específica e assim melhorar a eficiência e o direcionamento da edição gênica. Nesse trabalho utilizamos o silenciamento gênico mediano por short interference RNA (siRNA) para inibição temporária dos genes ATF7IP uma metiltrasferase, EP300 uma acetiltransferase e KU70 (NHEJ) e um par de TALENs complementares a uma região do gene da distrofina canina. Células Caninas MDCK I foram transfectadas por lipofectamina 2000 (Invitrogen) com 320pmol de siRNAs para ATF7IP e Ep300; e 64 pmol do SiRNA para KU70 em diferentes grupos, 40 horas depois as células foram transfectadas com 15 μg vetor molde derivado do pEGFP-N1 (Clonatech) e com 10 μg dos RNAm das TALENs. A seleção se deu em meio DMEM high com 600μg/ mL de G418 (Lonza) por 14-16 dias. As colônias coletadas através de biópsias foram analisadas por Polimerase Chain Reaction e sequenciamento gênico. Três pares de primers foram utilizados; um controle endógeno (GAPDH), um controle interno do inserto (Neo qPCR) e um para confirmação da recombinação homóloga (DMD3). Os grupos apresentaram grande variação na taxa de mortalidade celular e consequentemente no número de colônias: Com o grupo ATF7IP+Vetor (648c) apresentando maior número de colônias e o grupo EP300+Ku70+Vetor+TALENs o menor (1c). A maior taxa de recombinação ocorreu nos grupos no grupo ATF7IP +Ku70+Vetor+TALENs com 40% das células positivas para neomicina apresentado o evento gene targeting, um aumento considerável na taxa de recombinação quando comparada a porcentagem de 3,1% do controle transfectado somente com o vetor molde. Mostrando que o uso conjunto das TALENs com siRNAs foi um sucesso para o aumento de eventos de edição gênica direcionada.

The insertion of exogenous DNA into a host genome is achieved primarily through the use of DNA repair pathways such as Non-Homologous End Joining (NHEJ) and the Homologous Recombination (HR). The integration by NHEJ has a random feature and is much more common than HR insertions, which are more likely to produce gene targeting events . TAL effector nucleases (TALENs) and RNA interference (RNAi) can be used to increase the rate of specific integration and thus improving the efficiency of gene editing. In this work, we used short interference RNA (siRNA)-mediated gene silencing for transient inhibition of genes ATF7IP (implicated in histone methylation), EP300 (acetyltransferase) and Ku70 (essential to NHEJ) and a pair of TALENs RNAm complementary to canine muscle dystrophin (DMD) gene. MDCK I Canine Cells were transfected by lipofectamine 2000 (Invitrogen) with 320 pmol of siRNAs for ATF7IP and EP300; and 64 pmol of siRNA for Ku70 in different groups. After 40 hours cells were…

Advisors/Committee Members: Ambrosio, Carlos Eduardo.

Subjects/Keywords: Gene targeting; Gene targeting; Homologous recombination; Recombinação homóloga; RNAi; RNAi; TALENs; TALENs

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Pinho, R. d. M. e. (2014). Uso do silenciamento gênico mediado por RNA de interferência e de TAL effector nucleases para aumento de eventos gene targeting em células de cão. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/10/10132/tde-13012015-102643/ ;

Chicago Manual of Style (16^th Edition):

Pinho, Raquel de Mello e. “Uso do silenciamento gênico mediado por RNA de interferência e de TAL effector nucleases para aumento de eventos gene targeting em células de cão.” 2014. Masters Thesis, University of São Paulo. Accessed November 18, 2019. http://www.teses.usp.br/teses/disponiveis/10/10132/tde-13012015-102643/ ;.

MLA Handbook (7^th Edition):

Pinho, Raquel de Mello e. “Uso do silenciamento gênico mediado por RNA de interferência e de TAL effector nucleases para aumento de eventos gene targeting em células de cão.” 2014. Web. 18 Nov 2019.

Vancouver:

Pinho RdMe. Uso do silenciamento gênico mediado por RNA de interferência e de TAL effector nucleases para aumento de eventos gene targeting em células de cão. [Internet] [Masters thesis]. University of São Paulo; 2014. [cited 2019 Nov 18]. Available from: http://www.teses.usp.br/teses/disponiveis/10/10132/tde-13012015-102643/ ;.

Council of Science Editors:

Pinho RdMe. Uso do silenciamento gênico mediado por RNA de interferência e de TAL effector nucleases para aumento de eventos gene targeting em células de cão. [Masters Thesis]. University of São Paulo; 2014. Available from: http://www.teses.usp.br/teses/disponiveis/10/10132/tde-13012015-102643/ ;

University of Toronto

14. Seigel, Kyle. Enhancing the Efficiency of CRISPR/Cas9 Precise Gene Editing for Cystic Fibrosis Gene Therapy.

Degree: 2018, University of Toronto

URL: http://hdl.handle.net/1807/91348

►

Cystic fibrosis (CF) is the most common cause of chronic obstructive lung disease in children and young adults, yet there is no cure for this… (more)

Subjects/Keywords: CRISPR/Cas9; CtIP; Cystic fibrosis; EXO1; Gene targeting; Gene therapy; 0564

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Seigel, K. (2018). Enhancing the Efficiency of CRISPR/Cas9 Precise Gene Editing for Cystic Fibrosis Gene Therapy. (Masters Thesis). University of Toronto. Retrieved from http://hdl.handle.net/1807/91348

Chicago Manual of Style (16^th Edition):

Seigel, Kyle. “Enhancing the Efficiency of CRISPR/Cas9 Precise Gene Editing for Cystic Fibrosis Gene Therapy.” 2018. Masters Thesis, University of Toronto. Accessed November 18, 2019. http://hdl.handle.net/1807/91348.

MLA Handbook (7^th Edition):

Seigel, Kyle. “Enhancing the Efficiency of CRISPR/Cas9 Precise Gene Editing for Cystic Fibrosis Gene Therapy.” 2018. Web. 18 Nov 2019.

Vancouver:

Seigel K. Enhancing the Efficiency of CRISPR/Cas9 Precise Gene Editing for Cystic Fibrosis Gene Therapy. [Internet] [Masters thesis]. University of Toronto; 2018. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/1807/91348.

Council of Science Editors:

Seigel K. Enhancing the Efficiency of CRISPR/Cas9 Precise Gene Editing for Cystic Fibrosis Gene Therapy. [Masters Thesis]. University of Toronto; 2018. Available from: http://hdl.handle.net/1807/91348

University of Southern California

15. Lei, Yuning. Engineering viral vectors for gene and cell targeting.

Degree: PhD, Chemical Engineering, 2011, University of Southern California

URL: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/430755/rec/2369

► Gene therapy is the introduction of functional genes into dysfunctional cells to treat potentially incurable diseases. The technique sounds promising, yet there are many hurdles… (more)

▼ Gene therapy is the introduction of functional genes into dysfunctional cells to treat potentially incurable diseases. The technique sounds promising, yet there are many hurdles must be overcome to make it a practical mean of medicine. Viral vector mediated gene therapy remains one of the most promising gene therapy techniques as its efficiency and duration is the highest among the different gene delivery vehicles. To further refine the viral vector to enhance the gene delivery ability, we designed a strategy by decoupling binding and fusion ability of envelope protein into two distinct proteins. By pseudotyping the viral vectors with both an antibody and a fusogenic molecule, we can target and transduce specific cell types. The underlining mechanism of targeted transduction is that the viral vector will bind to the desirable cell types via the cognate antibody antigen interaction and further be endocytosised into the endosome. Inside the endosome compartment, the low pH environment will trigger the conformation change of the fusogenic molecule which will fuse both the viral and cellular membrane, resulting in releasing the viral core. To further enhance the targeted transduction, we focused on engineering the two parameters on the viral vector. In chapter 2, we engineered the fusion loop of the fusogenic molecule to enhance the activity of the protein. We demonstrated that the engineered fusogenic molecules can enhance targeted transduction many folds. We further looked at the other parameter, the targeting antibody in chapter 3. By designing a single chain antibody, we were able to constructed our targeting virus a simplifier viral production protocol and showed that the viral vector still exhibit targeted transduction. These two chapters concluded our effort in enhancing the targeted transduction by genetically engineering the component displayed on the viral membrane.; The fourth chapter in this report combines the technology of the zinc finger nuclease and the baculoviral vector. Currently, one of the drawback of utilizing integrating viral vector is the possibility of turning on oncogenes when the viral vectors integrated to undesirable sites. In this chapter, we utilize zinc finger nuclease to generate double stranded break and further demonstrate the ability of utilizing this technique to converting the baculoviral vector into gene targeting vector. We showed that our system can targeted transduce both immortalized cell lines as well as human embryonic stem cells. Techniques that we developed here can further enhance the safety of viral gene therapy while retain the efficiency and duration of the viral vectors. Advisors/Committee Members: Wang, Pin (Committee Chair), Shing, Katherine S. (Committee Member), Arnold, Donald B. (Committee Member).

Subjects/Keywords: lentiviral vector; baculoviral vector; CD20 targeting; cell targeting; zinc finger nuclease; gene targeting; human embryonic stem cells; genetic modification

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Lei, Y. (2011). Engineering viral vectors for gene and cell targeting. (Doctoral Dissertation). University of Southern California. Retrieved from http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/430755/rec/2369

Chicago Manual of Style (16^th Edition):

Lei, Yuning. “Engineering viral vectors for gene and cell targeting.” 2011. Doctoral Dissertation, University of Southern California. Accessed November 18, 2019. http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/430755/rec/2369.

MLA Handbook (7^th Edition):

Lei, Yuning. “Engineering viral vectors for gene and cell targeting.” 2011. Web. 18 Nov 2019.

Vancouver:

Lei Y. Engineering viral vectors for gene and cell targeting. [Internet] [Doctoral dissertation]. University of Southern California; 2011. [cited 2019 Nov 18]. Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/430755/rec/2369.

Council of Science Editors:

Lei Y. Engineering viral vectors for gene and cell targeting. [Doctoral Dissertation]. University of Southern California; 2011. Available from: http://digitallibrary.usc.edu/cdm/compoundobject/collection/p15799coll127/id/430755/rec/2369

16. Merle, Jacob Andrew. Evidence for IRES Mediated Translation of Gurken.

Degree: 2014, SUNY College at Fredonia

URL: http://hdl.handle.net/1951/65149

► The gene gurken (grk) in Drosophila melanogaster is required to establish the anterior/posterior and dorsal/ventral axes of the egg. When insulin levels are high such… (more)

▼ The gene gurken (grk) in Drosophila melanogaster is required to establish the anterior/posterior and dorsal/ventral axes of the egg. When insulin levels are high such as when food is readily available, grk is translated using a common mechanism that requires recognition of the 7-methylguanosine cap at the 5’ end of the RNA. Translation of grk requires Vasa activity which is inhibited through phosphorylation is spindle-B (spn-B) mutants. It was discovered in our lab that Insulin/Insulin-like Signaling (IIS) mutations or inhibition of TOR by rapamycin result in increased grk translation in spn-B mutants thereby overcoming this cap-dependent block. Based on these data, we hypothesize that the grk 5’ UTR contains an Internal Ribosomal Entry Site (IRES). An IRES provides an alternative mechanism of translation that occurs through use of a secondary structure in the 5’ UTR in the mRNA. The utility of the IRES is that it allows for translation of important mRNA’s even in the absence of canonical cap-dependent translation factors. These conditions can arise in nature when insulin levels or nutrient availability is low. The presence of an IRES will be explored in vivo through a comparison of transgenic reporter lines containing different luciferase reporter constructs. These constructs will be used to test the hypothesis that the grk 5’ UTR contains an IRES and examine the effect of nutrient limitation, inhibition of TOR, or IIS mutations on grk translation. To localize the IRES within the GRK 5' UTR selective deletion mutations will be made in secondary structures identified through selective 2’-Hydroxyl Acylation and Primer Extension (SHAPE) analysis. The activity of these reporter lines will be analyzed through assaying these transgenic lines. This will allow for the identification of the presence and location of the IRES within the grk 5’ UTR. Here we present data demonstrating the activity of reporter constructs that have been generated by fusion with the 5' UTR of grk. Additionally we propose a new technique using GRNA chromatography to identify the presence of an IRES as well as important secondary structures needed for IRES function using the same reporter constructs.

Subjects/Keywords: Drosophila melanogaster.; Nucleotide sequence.; Biology – Experiments.; Gene targeting.

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APA (6^th Edition):

Merle, J. A. (2014). Evidence for IRES Mediated Translation of Gurken. (Masters Thesis). SUNY College at Fredonia. Retrieved from http://hdl.handle.net/1951/65149

Chicago Manual of Style (16^th Edition):

Merle, Jacob Andrew. “Evidence for IRES Mediated Translation of Gurken. ” 2014. Masters Thesis, SUNY College at Fredonia. Accessed November 18, 2019. http://hdl.handle.net/1951/65149.

MLA Handbook (7^th Edition):

Merle, Jacob Andrew. “Evidence for IRES Mediated Translation of Gurken. ” 2014. Web. 18 Nov 2019.

Vancouver:

Merle JA. Evidence for IRES Mediated Translation of Gurken. [Internet] [Masters thesis]. SUNY College at Fredonia; 2014. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/1951/65149.

Council of Science Editors:

Merle JA. Evidence for IRES Mediated Translation of Gurken. [Masters Thesis]. SUNY College at Fredonia; 2014. Available from: http://hdl.handle.net/1951/65149

University of Manchester

17. Costello, Ryan. Towards the analyses of cell lineages using conditional gene alterations.

Degree: PhD, 2016, University of Manchester

URL: https://www.research.manchester.ac.uk/portal/en/theses/towards-the-analyses-of-cell-lineages-using-conditional-gene-alterations(af6e20c3-1dd5-4c46-b14e-e062900b812a).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.684836

► The ability to precisely modify the mouse genome is an invaluable tool for any researcher. If an artificial epitope sequence is integrated into target loci… (more)

▼ The ability to precisely modify the mouse genome is an invaluable tool for any researcher. If an artificial epitope sequence is integrated into target loci in specific cell types, it is possible to generate mice with these cells specifically tagged with the epitope, which can be used for many subsequent studies. Homologous recombination and the Cre/loxP system have been used to generate targeted and conditional transgenic mice, which have provided the basis for many studies into gene function. However, in recent years, improvements in technology have led to the development of RNA and protein based methods of specifically editing DNA sequences at user-selected loci. This thesis aimed to investigate the effectiveness of the novel gene targeting methods TALEN and CRISPR/Cas9. It also aimed to utilize different strains of mice generated using the Cre/loxP system in Trichuris muris, an animal infection model of the human disease Trichuris trichiura. TALENs use a pair of protein-based monomers specific to the sense and anti-sense strand of a target DNA sequence to dimerise a FokI nuclease and initiate a deletion in the genome. As a study into the practical use of this emerging technology TALENs were generated to target Oct-4 (a stem cell marker) in order to integrate an artificial epitope sequence, which could be used for enrichment experiments. The CRISPR/Cas9 is a very efficient RNA-based system used for modifying a target sequence. This system has been utilized to integrate an epitope sequence into the Rosa26 locus, downstream of a floxed STOP codon. This allows for expression of the epitope following the introduction of tissue restricted Cre recombinase. IL-1 is an important cytokine in the immune response towards T.muris. IL-1R1 was conditionally removed in CD4 cells and the role of IL-1 signaling in developing Th1, Th2 and Th17 responses was then studied. Interestingly, IL-1R1fl/fl CD4Cre mice could generate Th1 and Th2 response but showed a reduction in IL-22 and IL-17 production, two key Th17 cytokines. Infected IL-1R1fl/fl CD4Cre mice also displayed increased gut morphology and goblet cell hyperplasia. Therefore, it was concluded that IL-1 signaling from CD4 cells has an important role in host defense and the development of a full Th17 response. It was also shown that removing IL-1R1 in naïve mice had no affect on lymphocyte development. IL-10 is an anti-inflammatory cytokine expressed by gut macrophages, which contributes to homeostatic control of the immune system. IL-10R was specifically removed in the macrophage specific Cre lines LysMCre and also in CX3CR1Cre as a way of comparing the two Cre drivers. The mice were then infected with T.muris and displayed significant inflammation and also failure to expel the worms in the LysMCre model. This suggests a role for this model in future studies of gut macrophages. Clearly, animal models are very important in the study of gene function and also as a method of assessing the application of new technologies such as CRISPR/Cas9. Future work with the artificial epitope…

Subjects/Keywords: 572.8; Gene Targeting; CRISPR; TALEN; IL-1; CD4; Rosa26; Artificial Epitope

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Costello, R. (2016). Towards the analyses of cell lineages using conditional gene alterations. (Doctoral Dissertation). University of Manchester. Retrieved from https://www.research.manchester.ac.uk/portal/en/theses/towards-the-analyses-of-cell-lineages-using-conditional-gene-alterations(af6e20c3-1dd5-4c46-b14e-e062900b812a).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.684836

Chicago Manual of Style (16^th Edition):

Costello, Ryan. “Towards the analyses of cell lineages using conditional gene alterations.” 2016. Doctoral Dissertation, University of Manchester. Accessed November 18, 2019. https://www.research.manchester.ac.uk/portal/en/theses/towards-the-analyses-of-cell-lineages-using-conditional-gene-alterations(af6e20c3-1dd5-4c46-b14e-e062900b812a).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.684836.

MLA Handbook (7^th Edition):

Costello, Ryan. “Towards the analyses of cell lineages using conditional gene alterations.” 2016. Web. 18 Nov 2019.

Vancouver:

Costello R. Towards the analyses of cell lineages using conditional gene alterations. [Internet] [Doctoral dissertation]. University of Manchester; 2016. [cited 2019 Nov 18]. Available from: https://www.research.manchester.ac.uk/portal/en/theses/towards-the-analyses-of-cell-lineages-using-conditional-gene-alterations(af6e20c3-1dd5-4c46-b14e-e062900b812a).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.684836.

Council of Science Editors:

Costello R. Towards the analyses of cell lineages using conditional gene alterations. [Doctoral Dissertation]. University of Manchester; 2016. Available from: https://www.research.manchester.ac.uk/portal/en/theses/towards-the-analyses-of-cell-lineages-using-conditional-gene-alterations(af6e20c3-1dd5-4c46-b14e-e062900b812a).html ; http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.684836

18. Novo, Luís. Decationized polyplexes for targeted delivery of nucleic acids : from carrier design to in vivo evaluation.

Degree: 2014, Universiteit Utrecht

URL: http://dspace.library.uu.nl:8080/handle/1874/300546

► Gene therapy is considered a promising treatment for current intractable diseases. However, the clinical applicability of gene therapy is highly dependent on the development of… (more)

▼ Gene therapy is considered a promising treatment for current intractable diseases. However, the clinical applicability of gene therapy is highly dependent on the development of safe and efficient gene delivery vectors. So far, viral vectors have been used for clinical applications, but due to severe risks associated with viruses, cationic polymers have been evaluated as alternatives to viral vectors. Cationic polymers can easily form nanosized particles with plasmid DNA (pDNA), named polyplexes, via electrostatic interactions and possess an enormous chemical and structural flexibility. Polycation-based vectors have demonstrated high efficiency in vitro, however, they induce severe toxicity and possess suboptimal efficiencies in vivo, mainly due to their cationic nature, which significantly hampers their clinical applicability. With our work we have developed an alternative to conventional polycation-based polyplexes: decationized polyplexes. Unlike the cationic polymer based systems, decationized polyplexes are formed by hydrophilic and neutral polymers and can be obtained by an innovative 3-step process: polyplex formation by electrostatic interaction between pDNA and a polycationic precursor, structure stabilization by disulfide crosslinking, and finally removal of cationic charge - decationization. Structurally, decationized polyplexes consist of a disulfide-crosslinked poly(hydroxypropyl methacrylamide) (pHPMA) core stably entrapping plasmid DNA (pDNA), surrounded by a shell of poly(ethylene glycol) (PEG). Retention of pDNA in the nanoparticles is exclusively based on physical entrapment given by the disulfide crosslinks, which provides an intracellularly triggered release profile, since disulfides are cleaved under the higher reducing environment present inside the cells. Through our study decationized polyplexes have demonstrated important advantages when compared to their cationic counterparts, such as much lower degree of nonspecific uptake and high degree cell specific uptake when decorated with targeting moieties as demonstrated by several cell uptake studies. Furthermore, in vitro studies showed lower cellular toxicity and in vivo nanotoxicity studies using a zebrafish model showed remarkable lower teratogenicity and mortality profile from decationized polyplexes. Stability evaluation in biological fluids a high stability for prolonged periods was found. Finally, in an in vivo biodistribution study, using tumor bearing mice, decationized polyplexes have shown greater retention in blood circulation and higher target tissue (tumor) accumulation. Given their important advantages, decationized polyplexes were also investigated and optimized for small interfering RNA (siRNA) delivery purposes, showing that decationized polyplexes can be used as a platform for different gene delivery modalities. In conclusion, decationzed polyplexes have demonstrated to be an important contribution for the development of safer polymeric gene delivery systems especially for targeted therapies. Importantly, the requirements for… Advisors/Committee Members: Hennink, Wim, van Nostrum, Rene, Mastrobattista, Enrico.

Subjects/Keywords: Gene delivery; pDNA; siRNA; polymer; biocompatibility; nanoparticle; targeting; biodistribution; in vivo

11 more images…

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Novo, L. (2014). Decationized polyplexes for targeted delivery of nucleic acids : from carrier design to in vivo evaluation. (Doctoral Dissertation). Universiteit Utrecht. Retrieved from http://dspace.library.uu.nl:8080/handle/1874/300546

Chicago Manual of Style (16^th Edition):

Novo, Luís. “Decationized polyplexes for targeted delivery of nucleic acids : from carrier design to in vivo evaluation.” 2014. Doctoral Dissertation, Universiteit Utrecht. Accessed November 18, 2019. http://dspace.library.uu.nl:8080/handle/1874/300546.

MLA Handbook (7^th Edition):

Novo, Luís. “Decationized polyplexes for targeted delivery of nucleic acids : from carrier design to in vivo evaluation.” 2014. Web. 18 Nov 2019.

Vancouver:

Novo L. Decationized polyplexes for targeted delivery of nucleic acids : from carrier design to in vivo evaluation. [Internet] [Doctoral dissertation]. Universiteit Utrecht; 2014. [cited 2019 Nov 18]. Available from: http://dspace.library.uu.nl:8080/handle/1874/300546.

Council of Science Editors:

Novo L. Decationized polyplexes for targeted delivery of nucleic acids : from carrier design to in vivo evaluation. [Doctoral Dissertation]. Universiteit Utrecht; 2014. Available from: http://dspace.library.uu.nl:8080/handle/1874/300546

University of Manchester

19. Costello, Ryan. Towards the analyses of cell lineages using conditional gene alterations.

Degree: 2016, University of Manchester

URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:295855

► The ability to precisely modify the mouse genome is an invaluable tool for any researcher. If an artificial epitope sequence is integrated into target loci… (more)

▼ The ability to precisely modify the mouse genome is an invaluable tool for any researcher. If an artificial epitope sequence is integrated into target loci in specific cell types, it is possible to generate mice with these cells specifically tagged with the epitope, which can be used for many subsequent studies. Homologous recombination and the Cre/loxP system have been used to generate targeted and conditional transgenic mice, which have provided the basis for many studies into gene function. However, in recent years, improvements in technology have led to the development of RNA and protein based methods of specifically editing DNA sequences at user-selected loci. This thesis aimed to investigate the effectiveness of the novel gene targeting methods TALEN and CRISPR/Cas9. It also aimed to utilize different strains of mice generated using the Cre/loxP system in Trichuris muris, an animal infection model of the human disease Trichuris trichiura. TALENs use a pair of protein-based monomers specific to the sense and anti-sense strand of a target DNA sequence to dimerise a FokI nuclease and initiate a deletion in the genome. As a study into the practical use of this emerging technology TALENs were generated to target Oct-4 (a stem cell marker) in order to integrate an artificial epitope sequence, which could be used for enrichment experiments. The CRISPR/Cas9 is a very efficient RNA-based system used for modifying a target sequence. This system has been utilized to integrate an epitope sequence into the Rosa26 locus, downstream of a floxed STOP codon. This allows for expression of the epitope following the introduction of tissue restricted Cre recombinase.IL-1 is an important cytokine in the immune response towards T.muris. IL-1R1 was conditionally removed in CD4 cells and the role of IL-1 signaling in developing Th1, Th2 and Th17 responses was then studied. Interestingly, IL-1R1fl/fl CD4Cre mice could generate Th1 and Th2 response but showed a reduction in IL-22 and IL-17 production, two key Th17 cytokines. Infected IL-1R1fl/fl CD4Cre mice also displayed increased gut morphology and goblet cell hyperplasia. Therefore, it was concluded that IL-1 signaling from CD4 cells has an important role in host defense and the development of a full Th17 response. It was also shown that removing IL-1R1 in naïve mice had no affect on lymphocyte development.IL-10 is an anti-inflammatory cytokine expressed by gut macrophages, which contributes to homeostatic control of the immune system. IL-10R was specifically removed in the macrophage specific Cre lines LysMCre and also in CX3CR1Cre as a way of comparing the two Cre drivers. The mice were then infected with T.muris and displayed significant inflammation and also failure to expel the worms in the LysMCre model. This suggests a role for this model in future studies of gut macrophages.Clearly, animal models are very important in the study of gene function and also as a method of assessing the application of new technologies such as CRISPR/Cas9. Future work with the artificial epitope… Advisors/Committee Members: TRAVIS, MARK MA, Muller, Werner, Travis, Mark.

Subjects/Keywords: Gene Targeting; CRISPR; TALEN; IL-1; CD4; Rosa26; Artificial Epitope

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Costello, R. (2016). Towards the analyses of cell lineages using conditional gene alterations. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:295855

Chicago Manual of Style (16^th Edition):

Costello, Ryan. “Towards the analyses of cell lineages using conditional gene alterations.” 2016. Doctoral Dissertation, University of Manchester. Accessed November 18, 2019. http://www.manchester.ac.uk/escholar/uk-ac-man-scw:295855.

MLA Handbook (7^th Edition):

Costello, Ryan. “Towards the analyses of cell lineages using conditional gene alterations.” 2016. Web. 18 Nov 2019.

Vancouver:

Costello R. Towards the analyses of cell lineages using conditional gene alterations. [Internet] [Doctoral dissertation]. University of Manchester; 2016. [cited 2019 Nov 18]. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:295855.

Council of Science Editors:

Costello R. Towards the analyses of cell lineages using conditional gene alterations. [Doctoral Dissertation]. University of Manchester; 2016. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:295855

Columbia University

20. SriRamaratnam, Rohitha. Application and development of methods towards the target identification of biologically-active small molecules.

Degree: 2011, Columbia University

URL: https://doi.org/10.7916/D8T169HJ

► Small molecules have played an important role in defining the functions and identities of numerous proteins involved in fundamental biological processes as well as pathways… (more)

▼ Small molecules have played an important role in defining the functions and identities of numerous proteins involved in fundamental biological processes as well as pathways involved in disease. Chemical genetics represents the formalization of this process into a defined field desiring to achieve the breadth and specificity of classical genetics. In order to gain full advantage of a small molecule's ability to perturb the cell for novel or desired phenotypes, a complete understanding of the molecule's mechanism of action must be achieved. Identification of the biological targets of a molecule represents the most direct approach to attaining this knowledge. In our strategy to find novel mechanisms to target cancers with oncogenic RAS mutations, we have used small molecules to probe specific weaknesses of this cancerous network through synthetic lethal screening. One molecule identified in these screens, RSL3, attracted interest as a candidate for target identification studies because of its potent lethality and potentially unique mechanism of action. We used an affinity chromatography approach to directly isolate binding partners of RSL3 by modifying the molecules structure to incorporate various affinity tags. Through these experiments we ultimately identified a number of interesting candidate targets. Investigations validating these targets suggest that multi-targeted modulation of antioxidant and prostaglandin networks may be a mechanism for selectively killing cancers with oncogenic RAS. The identification of biological targets of small molecules poses a difficult challenge to the field of forward chemical genetics. Thus, we attempted to optimize a unique method for target identification, the yeast three-hybrid system (Y3H), which detects small molecule-protein interactions through a transcriptional assay in vivo. We created a version of our Y3H system that incorporated a covalent anchor and compared it with the existing state-of-the-art, which uses a high affinity non-covalent anchor. Transcriptional assays indicated our new system was functional, but surprisingly could not improve upon the original Y3H system. These results highlight the complexities of manipulating ligand-receptor interactions in vivo.

Subjects/Keywords: Chemistry; Biology; Molecules; Biochemistry; Biochemical genetics; Gene targeting

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

SriRamaratnam, R. (2011). Application and development of methods towards the target identification of biologically-active small molecules. (Doctoral Dissertation). Columbia University. Retrieved from https://doi.org/10.7916/D8T169HJ

Chicago Manual of Style (16^th Edition):

SriRamaratnam, Rohitha. “Application and development of methods towards the target identification of biologically-active small molecules.” 2011. Doctoral Dissertation, Columbia University. Accessed November 18, 2019. https://doi.org/10.7916/D8T169HJ.

MLA Handbook (7^th Edition):

SriRamaratnam, Rohitha. “Application and development of methods towards the target identification of biologically-active small molecules.” 2011. Web. 18 Nov 2019.

Vancouver:

SriRamaratnam R. Application and development of methods towards the target identification of biologically-active small molecules. [Internet] [Doctoral dissertation]. Columbia University; 2011. [cited 2019 Nov 18]. Available from: https://doi.org/10.7916/D8T169HJ.

Council of Science Editors:

SriRamaratnam R. Application and development of methods towards the target identification of biologically-active small molecules. [Doctoral Dissertation]. Columbia University; 2011. Available from: https://doi.org/10.7916/D8T169HJ

University of New South Wales

21. Hook, Jeff. Functional identity of the Mammalian Gamma Tropomyosin Gene.

Degree: Medical Sciences, 2012, University of New South Wales

URL: http://handle.unsw.edu.au/1959.4/52274 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:10946/SOURCE01?view=true

► The actin filament system is fundamental to cellular functions including regulation of shape, motility, cytokinesis, intracellular trafficking and tissue organisation. Tropomyosins (Tm) are highly conserved… (more)

▼ The actin filament system is fundamental to cellular functions including regulation of shape, motility, cytokinesis, intracellular trafficking and tissue organisation. Tropomyosins (Tm) are highly conserved components of actin filaments which differentially regulate filament stability and function. The mammalian Tm family consists of four genes; alphaTm, betaTm, gammaTm and deltaTm (Tpm1-4). Multiple Tm isoforms (>40) are generated by alternative splicing, and the expression of these isoforms is highly regulated with spatial and temporal sorting of Tm isoforms into different cellular compartments. The importance of gammaTm gene products during mammalian development has not been fully explored. In order to further identify the role of mammalian Tm isoforms, the functional specificity of subsets of gammaTm gene family products in mice was tested using a series of gene knockouts. Knockout constructs of the amino-terminal exon 1b and carboxy-terminal exons 9a+9b, exon 9c and exon 9d from the gammaTm gene were used to assess the viability of ES cells and mice. The deletion of amino-terminal coding exon 1b ablates all eleven known gammaTm gene cytoskeletal products (Tm5NM1-11) and results in non-viable mice. However, the elimination of four exon 9c-containing isoforms (Tm5NM4,7,8,9) shows no impact on embryo viability whereas the deletion of two exon 9d-containing isoforms (Tm5NM1,2) results in partial lethality. The viability of knockout ES cells in vitro was also compromised as neither exon 1b nor exon 9d homozygous knockout ES cells could be generated. In contrast, homozygous knockout ES cells for exons 9a+9b (Tm5NM3,5,6,8,9,10,11) were viable. Results indicate that exons 9a+9b may be required for ES cells to undergo differentiation and a conditional knockout mouse model of exons 9a+9b is being generated to determine whether these gene products are required for embryogenesis. Furthermore, sperm lacking cytoskeletal Tm products of the gammaTm gene show preferential transmission of the deleted allele which may indicate a selective advantage. Since all four Tm genes are expressed in early embryos, ES cells and sperm, it is concluded that isoforms from the gammaTm gene perform specific functions in ES cell viability and embryogenesis that cannot be compensated by the other Tm genes. Advisors/Committee Members: Gunning, Peter, Medical Sciences, Faculty of Medicine, UNSW, Kavallaris, Maria, Children's Cancer Institute Australia for Medical Research, Faculty of Medicine, UNSW.

Subjects/Keywords: Knockout mice; Cytoskeleton; Tropomyosin; Gene targeting; Embryo development

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Hook, J. (2012). Functional identity of the Mammalian Gamma Tropomyosin Gene. (Doctoral Dissertation). University of New South Wales. Retrieved from http://handle.unsw.edu.au/1959.4/52274 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:10946/SOURCE01?view=true

Chicago Manual of Style (16^th Edition):

Hook, Jeff. “Functional identity of the Mammalian Gamma Tropomyosin Gene.” 2012. Doctoral Dissertation, University of New South Wales. Accessed November 18, 2019. http://handle.unsw.edu.au/1959.4/52274 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:10946/SOURCE01?view=true.

MLA Handbook (7^th Edition):

Hook, Jeff. “Functional identity of the Mammalian Gamma Tropomyosin Gene.” 2012. Web. 18 Nov 2019.

Vancouver:

Hook J. Functional identity of the Mammalian Gamma Tropomyosin Gene. [Internet] [Doctoral dissertation]. University of New South Wales; 2012. [cited 2019 Nov 18]. Available from: http://handle.unsw.edu.au/1959.4/52274 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:10946/SOURCE01?view=true.

Council of Science Editors:

Hook J. Functional identity of the Mammalian Gamma Tropomyosin Gene. [Doctoral Dissertation]. University of New South Wales; 2012. Available from: http://handle.unsw.edu.au/1959.4/52274 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:10946/SOURCE01?view=true

University of Minnesota

22. Levine, Rachel. Exploring Critical Vehicle Parameters for the Design of Multitargeted Nanoparticles for Cancer Specific Gene Delivery.

Degree: PhD, Chemical Engineering, 2015, University of Minnesota

URL: http://hdl.handle.net/11299/188940

► The greatest obstacle to clinical application of cancer gene therapy is lack of effective delivery tools. Gene delivery vehicles must protect against degradation, avoid immunogenic… (more)

▼ The greatest obstacle to clinical application of cancer gene therapy is lack of effective delivery tools. Gene delivery vehicles must protect against degradation, avoid immunogenic effects and prevent off target delivery which can cause harmful side effects. The stealth liposomes has greatly improved tumor localization of small molecule drugs and is a promising tool for nucleic acid delivery as the polyethylene glycol coating its surface protects against immune recognition and blood clearance. In this study, DNA was fully encapsulated within in stealth liposomes by complexing the macromolecule with a cationic polymer before encapsulation. Formation methods and material compositions were then investigated for their effects on encapsulation. This technology was translated for protective delivery of siRNA designed for HPV viral gene silencing and cervical cancer treatment. Stealth liposomes encapsulating siRNA were functionalized with a targeting peptide which binds to the alpha6 integrin, a cervical cancer biomarker. It was found that both targeting and polymer complexation before encapsulation were critical components to effective transfection. Varying the siRNA:polymer ratio revealed an optimal concentration for enhanced transfection, but no improvement to internalization, suggesting polymer complexation enhances transfection through an intracellular mechanism. Nanoparticles functionalized with cancer-targeting ligands have shown promise but are still limited by off-tumor binding to healthy tissues that nevertheless express low levels of the molecular target. Targeting two unique cancer biomarkers using dual-targeted heteromultivalent nanoparticles presents a solution to this challenge by requiring overexpression of two separate ligands for localization. In order to guide experimental design, a kinetic model was built to explore how the affinity and valency of dual-ligand liposomes affect the binding and selectivity of delivery to cells with various receptor expression. α5β1 and α6β4 integrin expression levels were then quantified on 20 different cell lines to identify appropriate model cells for in vitro investigation. Dual-targeting heteromultivalent liposomes were synthesized using the alpha6 targeting peptide and an alpha5beta1 targeting peptide. Heteromultivalent liposomes with varying peptide ratios were delivered to cells with varying integrin concentrations. Binding and internalization was then evaluated to understand the effect of valency and avidity on binding kinetics and delivery. Dual-ligand liposomes with equal valencies of each targeting peptide achieved enhanced binding efficiency and selectivity for cells expressing equal and high receptor levels. This liposome formulation was used as a gene delivery vehicle to achieve improved transfection to dual-receptor expressing cells. The insights gained from this study inform rational design of modular heteromultivalent nanoparticles for enhanced specificity to target tissue, for the creation of more effective cancer treatments.

Subjects/Keywords: Dual Targeting; Gene Delivery; Peptide Amphiphile; Stealth Liposomes; Targeted Therapies

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Levine, R. (2015). Exploring Critical Vehicle Parameters for the Design of Multitargeted Nanoparticles for Cancer Specific Gene Delivery. (Doctoral Dissertation). University of Minnesota. Retrieved from http://hdl.handle.net/11299/188940

Chicago Manual of Style (16^th Edition):

Levine, Rachel. “Exploring Critical Vehicle Parameters for the Design of Multitargeted Nanoparticles for Cancer Specific Gene Delivery.” 2015. Doctoral Dissertation, University of Minnesota. Accessed November 18, 2019. http://hdl.handle.net/11299/188940.

MLA Handbook (7^th Edition):

Levine, Rachel. “Exploring Critical Vehicle Parameters for the Design of Multitargeted Nanoparticles for Cancer Specific Gene Delivery.” 2015. Web. 18 Nov 2019.

Vancouver:

Levine R. Exploring Critical Vehicle Parameters for the Design of Multitargeted Nanoparticles for Cancer Specific Gene Delivery. [Internet] [Doctoral dissertation]. University of Minnesota; 2015. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/11299/188940.

Council of Science Editors:

Levine R. Exploring Critical Vehicle Parameters for the Design of Multitargeted Nanoparticles for Cancer Specific Gene Delivery. [Doctoral Dissertation]. University of Minnesota; 2015. Available from: http://hdl.handle.net/11299/188940

University of Minnesota

23. Adil, Maroof Mohammad. A platform for next-generation cancer therapies: multi-targeted nonviral vectors for site-specific gene delivery and expression.

Degree: PhD, Chemical Engineering, 2013, University of Minnesota

URL: http://hdl.handle.net/11299/170877

► Advances in genetics have empowered gene therapy as a cancer treatment, however there are many challenges to delivering genes specifically to target disease sites. Presented… (more)

▼ Advances in genetics have empowered gene therapy as a cancer treatment, however there are many challenges to delivering genes specifically to target disease sites. Presented here is the development of a new non-viral gene delivery vehicle, consisting of branched polyethyleneimine (bPEI) condensed plasmid DNA polyplexes encapsulated within a PR_b functionalized stealth liposome, for the delivery of genes specifically to &alpha₅&beta₁ integrin overexpressing cancer cells. This new transfection agent mediated higher gene expression than non-targeted stealth liposomes and unencapsulated polyplexes in tissue culture. In a liver-metastatic colorectal cancer mouse model, PR_b functionalized stealth liposomes outperformed non-targeted stealth liposomes and was able to specifically transfect the tumor site while avoiding healthy tissues. In addition, a comparative investigation of the transfection mechanism of PR_b functionalized nanoparticles, DOTAP/DOPE lipoplexes, bPEI polyplexes and stealth liposomes was carried out in DLD-1 cells. Results demonstrated that PR_b functionalized nanoparticles were optimally balanced for the transfection of DLD-1 cells with high colloidal stability, fast integrin mediated internalization kinetics, caveolae mediated uptake and endosomal escape. To further increase the specificity of gene expression in cancer tissue, a new therapeutic plasmid DNA (pNF-&kappaB-DTA) was developed with expression of Diphtheria toxin fragment-A (DTA) gene regulated by the transcriptional activity of NF-&kappaB, which is a transcription factor upregulated in cancer. The multi-targeted gene delivery system formed by encapsulating pNF-&kappaB-DTA/bPEI polyplexes in PR_b functionalized stealth liposomes showed more specific gene expression in cancer cells versus healthy cells compared to either individually targeted system. Transfecting cancer cells using the multi-targeted gene delivery system resulted in a dose-dependent reduction of cellular protein expression and a dose-dependent increase in cytotoxicity. Our therapeutic delivery system specifically eradicated on average 70% of a variety of cancer cells while minimally affecting healthy cells. Moving forward, the modular nature of our non-viral delivery vehicle design can facilitate targeting novel pairs of extracellular receptors and upregulated transcription factors for applications beyond cancer gene therapy.

Subjects/Keywords: Cancer; Gene delivery; Integrin; Liposomes; NF-kB; Transcriptional targeting; Chemical engineering

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Adil, M. M. (2013). A platform for next-generation cancer therapies: multi-targeted nonviral vectors for site-specific gene delivery and expression. (Doctoral Dissertation). University of Minnesota. Retrieved from http://hdl.handle.net/11299/170877

Chicago Manual of Style (16^th Edition):

Adil, Maroof Mohammad. “A platform for next-generation cancer therapies: multi-targeted nonviral vectors for site-specific gene delivery and expression.” 2013. Doctoral Dissertation, University of Minnesota. Accessed November 18, 2019. http://hdl.handle.net/11299/170877.

MLA Handbook (7^th Edition):

Adil, Maroof Mohammad. “A platform for next-generation cancer therapies: multi-targeted nonviral vectors for site-specific gene delivery and expression.” 2013. Web. 18 Nov 2019.

Vancouver:

Adil MM. A platform for next-generation cancer therapies: multi-targeted nonviral vectors for site-specific gene delivery and expression. [Internet] [Doctoral dissertation]. University of Minnesota; 2013. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/11299/170877.

Council of Science Editors:

Adil MM. A platform for next-generation cancer therapies: multi-targeted nonviral vectors for site-specific gene delivery and expression. [Doctoral Dissertation]. University of Minnesota; 2013. Available from: http://hdl.handle.net/11299/170877

University of Texas Southwestern Medical Center

24. Borkowski, Robert John. Identifying Context-Specific Synthetic Lethal miRNA Inhibition in Non-Small Cell Lung Cancer.

Degree: 2013, University of Texas Southwestern Medical Center

URL: http://hdl.handle.net/2152.5/1726

► Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related fatalities in the US. This is due in part to a lack of highly… (more)

▼ Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related fatalities in the US. This is due in part to a lack of highly effective therapies for advanced cases, and this is of special concern as most NSCLC cases are not diagnosed until they are in an advanced, later stage. Recent successes in developing genotypically-targeted therapies with potency only in a well-defined subpopulation of tumors suggests that identifying targeted therapies for additional common NSCLC genotypes will improve patient survival. In this study I utilized a library of inhibitors to microRNAs, a class of post-transcriptional gene regulators, to identify novel synthetic lethal miRNA inhibition:molecular mechanism interactions in NSCLC. I accomplished this by screening a panel of 13 NSCLC and immortalized normal lung epithelium (HBEC) cell lines in two phases to identify miRNA inhibitors with selective toxicity in the NSCLC cell lines that were also benign in an HBEC cell line. Two inhibitors, the miR-92a and miR-1226* inhibitors, met these criteria. I then collected toxicity data in an expanded panel of 29 total cell lines. This expanded toxicity data was used to identify p53 loss as a molecular mechanism correlated with sensitivity to the miR-92a and miR-1226* inhibitors in NSCLC cell lines. This was recapitulated by demonstrating sensitivity after knockdown of p53 in the previously resistant HBEC30KT cell line. I determined that the inhibitors were toxic in a very sequence-specific manner and that they down-regulated the miR-17~92 polycistron. Down-regulation of the polycistron was toxic in a context-specific manner, and the down-regulation of the miR-17~92 cluster in sensitive cell lines mimicked activation of a 1α, 25-dihydroxyvitamin D3 response in NSCLC cell lines in a manner consistent with sensitivity to the miR-92a inhibitor. The results of this investigation demonstrate that the screening approach utilized in this study was capable of identifying a synthetic lethal miRNA inhibition:molecular mechanism interaction, and that I was then able to use a genetically defined model of the mechanism to identify a relevant mechanism of action for the toxic inhibitors. Advisors/Committee Members: Scaglioni, Pier Paolo, Pertsemlidis, Alexander, White, Michael A., Cobb, Melanie H., Corey, David R..

Subjects/Keywords: Carcinoma, Non-Small-Cell Lung; MicroRNAs; Gene Targeting

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APA (6^th Edition):

Borkowski, R. J. (2013). Identifying Context-Specific Synthetic Lethal miRNA Inhibition in Non-Small Cell Lung Cancer. (Thesis). University of Texas Southwestern Medical Center. Retrieved from http://hdl.handle.net/2152.5/1726

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Borkowski, Robert John. “Identifying Context-Specific Synthetic Lethal miRNA Inhibition in Non-Small Cell Lung Cancer.” 2013. Thesis, University of Texas Southwestern Medical Center. Accessed November 18, 2019. http://hdl.handle.net/2152.5/1726.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Borkowski, Robert John. “Identifying Context-Specific Synthetic Lethal miRNA Inhibition in Non-Small Cell Lung Cancer.” 2013. Web. 18 Nov 2019.

Vancouver:

Borkowski RJ. Identifying Context-Specific Synthetic Lethal miRNA Inhibition in Non-Small Cell Lung Cancer. [Internet] [Thesis]. University of Texas Southwestern Medical Center; 2013. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/2152.5/1726.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Borkowski RJ. Identifying Context-Specific Synthetic Lethal miRNA Inhibition in Non-Small Cell Lung Cancer. [Thesis]. University of Texas Southwestern Medical Center; 2013. Available from: http://hdl.handle.net/2152.5/1726

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Texas Southwestern Medical Center

25. Long, Chengzu. Prevention of Muscular Dystrophy in Mice by Gene Editing.

Degree: 2014, University of Texas Southwestern Medical Center

URL: http://hdl.handle.net/2152.5/3953

► Duchenne muscular dystrophy (DMD) is an inherited X-linked disease caused by mutations in the gene encoding dystrophin, a protein required for muscle fiber integrity. DMD… (more)

▼ Duchenne muscular dystrophy (DMD) is an inherited X-linked disease caused by mutations in the gene encoding dystrophin, a protein required for muscle fiber integrity. DMD is characterized by progressive muscle weakness and a shortened lifespan, often along with breathing and heart complications. There is no effective treatment. RNA-guided nucleases-mediated genome editing, based on Type II CRISPR/Cas systems, offers a new approach to alter the genome. It can precisely remove a mutation in DNA, allowing the DNA repair mechanisms to replace it with a normal copy of the gene. The benefit of this over other gene therapy techniques is that it can permanently correct the 'defect' in a gene rather than just transiently adding a 'functional' one. We used CRISPR/Cas9-mediated genome editing to correct the dystrophin gene (Dmd) mutation in the germline of mdx mice, a model for DMD, and then monitored skeletal muscle and heart structure and function. Genome editing produced genetically mosaic animals containing 2 to 100% correction of the Dmd gene. Histological analysis of skeletal muscle and heart from these corrected mice showed absence of the dystrophic muscle phenotype and restoration of dystrophin expression. In addition, the degree of muscle phenotypic rescue in mosaic mice exceeded the efficiency of gene correction, likely reflecting an advantage of the corrected stem cells and their contribution to regenerating muscle. Our experiments provide proof-of-concept that CRISPR/Cas9-mediated genomic editing can correct a causative germline mutation causing muscular dystrophy in a mouse model and prevent development of several characteristic features of the disease. With rapid technological advances of gene delivery systems and improvements to the CRISPR/Cas9 editing system, this strategy may allow correction of disease-causing mutations in the muscle tissue or iPSCs (induced pluripotent stem cells) from patients with genetic diseases. Advisors/Committee Members: Wang, Zhigao, Olson, Eric N., Cobb, Melanie H., Chen, Zhijian J..

Subjects/Keywords: CRISPR-Cas Systems; Dystrophin; Gene Targeting; Muscular Dystrophy, Duchenne

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APA (6^th Edition):

Long, C. (2014). Prevention of Muscular Dystrophy in Mice by Gene Editing. (Thesis). University of Texas Southwestern Medical Center. Retrieved from http://hdl.handle.net/2152.5/3953

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Long, Chengzu. “Prevention of Muscular Dystrophy in Mice by Gene Editing.” 2014. Thesis, University of Texas Southwestern Medical Center. Accessed November 18, 2019. http://hdl.handle.net/2152.5/3953.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Long, Chengzu. “Prevention of Muscular Dystrophy in Mice by Gene Editing.” 2014. Web. 18 Nov 2019.

Vancouver:

Long C. Prevention of Muscular Dystrophy in Mice by Gene Editing. [Internet] [Thesis]. University of Texas Southwestern Medical Center; 2014. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/2152.5/3953.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Long C. Prevention of Muscular Dystrophy in Mice by Gene Editing. [Thesis]. University of Texas Southwestern Medical Center; 2014. Available from: http://hdl.handle.net/2152.5/3953

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Texas Southwestern Medical Center

26. Voit, Richard Alexander 1983-. Generation of HIV-Resistant T-Cells and Correction of the Sickle Cell Mutation by Targeted Genome Engineering.

Degree: 2013, University of Texas Southwestern Medical Center

URL: http://hdl.handle.net/2152.5/3330

► Targeted genome engineering is a powerful method to create specific modifications at chromosomal loci. This technique makes it feasible to precisely alter DNA sequences by… (more)

▼ Targeted genome engineering is a powerful method to create specific modifications at chromosomal loci. This technique makes it feasible to precisely alter DNA sequences by introducing a specific DNA double-strand break, which is repaired by the natural cellular machinery. These double strand breaks are induced by engineered chimeric nucleases – either zinc finger nucleases (ZFNs) or Tal effector nucleases (TALENs) – and depending on the experimental design, can result in gene disruption, gene correction or targeted transgene integration. In this thesis, I present two applications of this approach in the context of two prevalent human diseases, HIV infection and sickle cell disease. HIV infects CD4+ T-cells by binding to the CD4 receptor and either the CCR5 or CXCR4 co-receptor on the surface of those cells. Previously, ZFNs were described that create gene specific knockouts of CCR5, protecting cells against CCR5-tropic (R5) HIV, but not against CXCR4-tropic (X4) HIV. I hypothesized that combining ZFN-mediated CCR5 disruption with targeted integration of a cassette of anti-HIV genes would confer higher levels of resistance against R5-tropic virus and also be protective against X4-tropic virus. In a T-cell reporter line, I showed that CCR5 disruption alone conferred 16-fold protection against R5-tropic virus but had no effect against X4-tropic HIV. In contrast, CCR5 disruption, combined with targeted gene integration into that locus, of the anti-HIV restriction factors human-rhesus hybrid TRIM5α, APOBEC3G D128K and Rev M10 was completely protective against both viral tropisms. Sickle cell disease is caused by a point mutation in the β-globin gene, and I sought to correct this mutation by synthesizing TALENs specific for that site. The β-globin TALENs stimulated integration of therapeutic β-globin cDNA in approximately 20% of cells prior to selection. Using FDA-approved drugs to select for modified cells, I showed virtually complete enrichment of targeted cells. Furthermore, I used the β-globin TALENs to target GFP to the β-globin start codon and designed TALENs to target tdTomato to the start codon of the γ-globin gene, upregulation of which is a goal of sickle cell disease pharmacotherapy. In this way, I developed an endogenous dual promoter reporter system and screened for drugs that preferentially upregulated γ-globin. Advisors/Committee Members: Goodman, Joel M., Porteus, Matthew H., Sternweis, Paul C., Graff, Jonathan M..

Subjects/Keywords: beta-Globins; Gene Targeting; HIV Infections; T-Lymphocytes

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APA (6^th Edition):

Voit, R. A. 1. (2013). Generation of HIV-Resistant T-Cells and Correction of the Sickle Cell Mutation by Targeted Genome Engineering. (Thesis). University of Texas Southwestern Medical Center. Retrieved from http://hdl.handle.net/2152.5/3330

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Voit, Richard Alexander 1983-. “Generation of HIV-Resistant T-Cells and Correction of the Sickle Cell Mutation by Targeted Genome Engineering.” 2013. Thesis, University of Texas Southwestern Medical Center. Accessed November 18, 2019. http://hdl.handle.net/2152.5/3330.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Voit, Richard Alexander 1983-. “Generation of HIV-Resistant T-Cells and Correction of the Sickle Cell Mutation by Targeted Genome Engineering.” 2013. Web. 18 Nov 2019.

Vancouver:

Voit RA1. Generation of HIV-Resistant T-Cells and Correction of the Sickle Cell Mutation by Targeted Genome Engineering. [Internet] [Thesis]. University of Texas Southwestern Medical Center; 2013. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/2152.5/3330.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Voit RA1. Generation of HIV-Resistant T-Cells and Correction of the Sickle Cell Mutation by Targeted Genome Engineering. [Thesis]. University of Texas Southwestern Medical Center; 2013. Available from: http://hdl.handle.net/2152.5/3330

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Guelph

27. Cealic, Iulia. Role of the Breast Cancer Susceptibility 2 BRC Repeats in Homologous Recombination .

Degree: 2013, University of Guelph

URL: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/5259

► Homologous recombination (HR) is a faithful mechanism for the repair of double-stranded DNA breaks (DSBs) and plays a critical role in maintaining the integrity of… (more)

Subjects/Keywords: BRCA2; BRC repeats; Homologous recombination; Gene targeting; Rad51

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APA (6^th Edition):

Cealic, I. (2013). Role of the Breast Cancer Susceptibility 2 BRC Repeats in Homologous Recombination . (Thesis). University of Guelph. Retrieved from https://atrium.lib.uoguelph.ca/xmlui/handle/10214/5259

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Cealic, Iulia. “Role of the Breast Cancer Susceptibility 2 BRC Repeats in Homologous Recombination .” 2013. Thesis, University of Guelph. Accessed November 18, 2019. https://atrium.lib.uoguelph.ca/xmlui/handle/10214/5259.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Cealic, Iulia. “Role of the Breast Cancer Susceptibility 2 BRC Repeats in Homologous Recombination .” 2013. Web. 18 Nov 2019.

Vancouver:

Cealic I. Role of the Breast Cancer Susceptibility 2 BRC Repeats in Homologous Recombination . [Internet] [Thesis]. University of Guelph; 2013. [cited 2019 Nov 18]. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/5259.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Cealic I. Role of the Breast Cancer Susceptibility 2 BRC Repeats in Homologous Recombination . [Thesis]. University of Guelph; 2013. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/5259

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Guelph

28. Knapp, Jennifer. Investigating the Role of Rad51 in Mammalian Ectopic Homologous Recombination.

Degree: 2013, University of Guelph

URL: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/7280

► DNA damage occurs through endogenous and exogenous sources, and can lead to stalled replication forks, genetic disorders, cancer, and cell death. Homologous recombination (HR) is… (more)

Subjects/Keywords: Ectopic homologous recombination; Murine hybridoma cells; Rad51; DNA damage; Gene Targeting

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APA (6^th Edition):

Knapp, J. (2013). Investigating the Role of Rad51 in Mammalian Ectopic Homologous Recombination. (Thesis). University of Guelph. Retrieved from https://atrium.lib.uoguelph.ca/xmlui/handle/10214/7280

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Knapp, Jennifer. “Investigating the Role of Rad51 in Mammalian Ectopic Homologous Recombination.” 2013. Thesis, University of Guelph. Accessed November 18, 2019. https://atrium.lib.uoguelph.ca/xmlui/handle/10214/7280.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Knapp, Jennifer. “Investigating the Role of Rad51 in Mammalian Ectopic Homologous Recombination.” 2013. Web. 18 Nov 2019.

Vancouver:

Knapp J. Investigating the Role of Rad51 in Mammalian Ectopic Homologous Recombination. [Internet] [Thesis]. University of Guelph; 2013. [cited 2019 Nov 18]. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/7280.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Knapp J. Investigating the Role of Rad51 in Mammalian Ectopic Homologous Recombination. [Thesis]. University of Guelph; 2013. Available from: https://atrium.lib.uoguelph.ca/xmlui/handle/10214/7280

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Florida Atlantic University

29. Helmick, Ericka Elizabeth. Genetic differentiation among populations of bald eagles, Haliaeetus leucocephalus.

Degree: MS, 2011, Florida Atlantic University

URL: http://purl.flvc.org/FAU/3171395

►

Summary: The bald eagle, Haliaeetus leucocephalus, population declined dramatically in the early 20th century reducing the population from tens of thousands of birds within the… (more)

Subjects/Keywords: Wildlife conservation; Birds – Molecular genetics; Gene targeting; Developmental biology; Biochemical markers

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Helmick, E. E. (2011). Genetic differentiation among populations of bald eagles, Haliaeetus leucocephalus. (Masters Thesis). Florida Atlantic University. Retrieved from http://purl.flvc.org/FAU/3171395

Chicago Manual of Style (16^th Edition):

Helmick, Ericka Elizabeth. “Genetic differentiation among populations of bald eagles, Haliaeetus leucocephalus.” 2011. Masters Thesis, Florida Atlantic University. Accessed November 18, 2019. http://purl.flvc.org/FAU/3171395.

MLA Handbook (7^th Edition):

Helmick, Ericka Elizabeth. “Genetic differentiation among populations of bald eagles, Haliaeetus leucocephalus.” 2011. Web. 18 Nov 2019.

Vancouver:

Helmick EE. Genetic differentiation among populations of bald eagles, Haliaeetus leucocephalus. [Internet] [Masters thesis]. Florida Atlantic University; 2011. [cited 2019 Nov 18]. Available from: http://purl.flvc.org/FAU/3171395.

Council of Science Editors:

Helmick EE. Genetic differentiation among populations of bald eagles, Haliaeetus leucocephalus. [Masters Thesis]. Florida Atlantic University; 2011. Available from: http://purl.flvc.org/FAU/3171395

30. Gras, Jan Cornelis Emile. A novel technology to target adenovirus vectors: application in cells involved in atherosclerosis.

Degree: 2007, Department of Human and Clinical Genetics, Medicine / Leiden University Medical Center (LUMC), Leiden University

URL: http://hdl.handle.net/1887/12431

► In this thesis a novel technology is described to target adenovirus vectors. Adenovirus vectors are powerful tools to modulate gene expression. The use of these… (more)

Subjects/Keywords: Adenovirus,,; Atherosclerosis; Gene Therapy; Targeting; Adenovirus,,; Atherosclerosis; Gene Therapy; Targeting

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APA (6^th Edition):

Gras, J. C. E. (2007). A novel technology to target adenovirus vectors: application in cells involved in atherosclerosis. (Doctoral Dissertation). Department of Human and Clinical Genetics, Medicine / Leiden University Medical Center (LUMC), Leiden University. Retrieved from http://hdl.handle.net/1887/12431

Chicago Manual of Style (16^th Edition):

Gras, Jan Cornelis Emile. “A novel technology to target adenovirus vectors: application in cells involved in atherosclerosis.” 2007. Doctoral Dissertation, Department of Human and Clinical Genetics, Medicine / Leiden University Medical Center (LUMC), Leiden University. Accessed November 18, 2019. http://hdl.handle.net/1887/12431.

MLA Handbook (7^th Edition):

Gras, Jan Cornelis Emile. “A novel technology to target adenovirus vectors: application in cells involved in atherosclerosis.” 2007. Web. 18 Nov 2019.

Vancouver:

Gras JCE. A novel technology to target adenovirus vectors: application in cells involved in atherosclerosis. [Internet] [Doctoral dissertation]. Department of Human and Clinical Genetics, Medicine / Leiden University Medical Center (LUMC), Leiden University; 2007. [cited 2019 Nov 18]. Available from: http://hdl.handle.net/1887/12431.

Council of Science Editors:

Gras JCE. A novel technology to target adenovirus vectors: application in cells involved in atherosclerosis. [Doctoral Dissertation]. Department of Human and Clinical Genetics, Medicine / Leiden University Medical Center (LUMC), Leiden University; 2007. Available from: http://hdl.handle.net/1887/12431

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Open Access Theses and Dissertations