subject:(RNA interference RNAi )

University of Edinburgh

1. Hall, Emma Andisi. Screening for genes involved in cilia formation and function.

Degree: PhD, 2012, University of Edinburgh

► Cilia are small microtubule based structures found on the surface of almost all mammalian cells, enclosed in a highly specialised extension of the cell membrane.… (more)

▼ Cilia are small microtubule based structures found on the surface of almost all mammalian cells, enclosed in a highly specialised extension of the cell membrane. Components of several key developmental signalling pathways, in particular Hedgehog (Hh) signalling, are enriched in cilia and cells with mutations in cilia structure show aberrant signalling, suggesting cilia act as “antennae” to focus these signalling cascades. A spectrum of human diseases, termed ciliopathies, are caused by problems in cilia formation or cilia function, which display wide ranging phenotypes from embryonic lethality to retinal degeneration, polydactyly to cystic kidneys. Despite recent advances in the understanding of the essential roles cilia play in mammalian development, exactly how these complex structures are put together, how they carry out their diverse functions, and how they are regulated is not well understood. In this thesis, I describe a screen for genes involved in cilia formation and function. While optimising ciliogenesis and immunofluorescence protocols for the screen, the phenotypes of two ciliary mutant cell lines were analysed. Wdr35yet/yet and Dync2h1pol/pol mouse lines were identified in an ENU screen for genes involved in early development, and shown to have gross phenotypes similar to other ciliary mutants (Mill et al. 2011). Intraflagellar transport (IFT) is the active transport of proteins up and down the ciliary axoneme. Dync2h1 is a retrograde IFT motor component, whereas Wdr35 is part of the retrograde IFT-A complex. In this thesis, the cellular phenotypes of mouse embryonic fibroblasts derived from these mutants are described, showing that despite the fact both genes are thought to be involved in retrograde IFT, they show distinct ciliary phenotypes, suggesting novel roles for Wdr35 in mouse ciliogenesis. An siRNA screen was carried out in mouse fibroblasts to identify genes involved in (i) cilia formation, assayed by immunofluorescence for ciliary markers, and (ii) cilia function, assayed by activity of a Hh responsive luciferase transgene as an indirect readout of ciliary function. Although scalable, I initially screened a small test set of thirty-six putative cilia candidates, identified by cross species transcriptomic analysis. We identified several possible hits, many of which were in the ciliome database but also importantly, several genes with no known link to ciliogenesis. Repeats, correlation of phenotype to knockdown efficiencies and localisation studies validated two hits, Ccdc63 and Azi1. Ccdc63 is a novel coiled-coil gene with no previous link to ciliogenesis; the phenotype for this gene was analysed in real time using fluorescently tagged ciliary markers. A second hit, Azi1, was followed up in more detail. The reduction in ciliogenesis upon Azi1 knockdown was confirmed with separate siRNAs, and was rescued by overexpressing siRNA insensitive Azi1-GFP, confirming the phenotype is not due to off-target effects of the siRNAs. Azi1 gene trap mutant mice were generated and confirmed to be null…

Subjects/Keywords: 572.8; cilia; Azi1; RNAi; RNA interference

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Hall, E. A. (2012). Screening for genes involved in cilia formation and function. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/9898

Chicago Manual of Style (16^th Edition):

Hall, Emma Andisi. “Screening for genes involved in cilia formation and function.” 2012. Doctoral Dissertation, University of Edinburgh. Accessed January 18, 2021. http://hdl.handle.net/1842/9898.

MLA Handbook (7^th Edition):

Hall, Emma Andisi. “Screening for genes involved in cilia formation and function.” 2012. Web. 18 Jan 2021.

Vancouver:

Hall EA. Screening for genes involved in cilia formation and function. [Internet] [Doctoral dissertation]. University of Edinburgh; 2012. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/1842/9898.

Council of Science Editors:

Hall EA. Screening for genes involved in cilia formation and function. [Doctoral Dissertation]. University of Edinburgh; 2012. Available from: http://hdl.handle.net/1842/9898

University of Iowa

2. Martin, Janine Nicole. Developing RNAi therapy For DYT1 dystonia.

Degree: PhD, Genetics, 2011, University of Iowa

URL: https://ir.uiowa.edu/etd/1019

► DYT1 dystonia is an early onset central nervous system-based movement disorder characterized by uncontrolled sustained muscle contractions that can lead to debilitating abnormal postures.… (more)

▼ DYT1 dystonia is an early onset central nervous system-based movement disorder characterized by uncontrolled sustained muscle contractions that can lead to debilitating abnormal postures. Though a genetic mutation in the gene TOR1A is responsible for most DYT1 cases, the low penetrance of the disease implicates additional genetic and environmental modifiers. Current therapeutic options for DYT1 dystonia are limited to symptomatic treatments with variable effectiveness. Currently, the underlying pathogenesis of this disease and the role of torsinA (torA), the protein product of TOR1A, in the development of this disease have yet to be established. In the first part of this thesis we aimed to further understand the effects of the TOR1A mutation at the molecular, cellular and organismal level in order to identify disease associated biomarkers that can be later used to measure the effectiveness of novel therapies. We found that expression of mutant torsinA (torA(ÄE)) in a cellular and an animal model of DYT1 had no significant effect on global transcription, despite its interaction with nuclear envelope proteins. Recent research has unearthed a role for microRNAs (miRNAs) in neuronal development and maturation. Consequently we explored whether torA(ÄE) expression in murine neural tissue was associated with changes in miRNA expression in young DYT1 knockin (KI) mice. Since the primary sight of dysfunction is still being debated, we profiled miRNA expression of the two strongest candidates, the striatum and cerebellum, both of which have well established roles in the control and coordination of muscle movements. We have identified several microRNAs that were uniquely altered in either the striatum or cerebellum and further research will be conducted to determine their usability as disease biomarkers. Finally, we were unable to identify motor phenotypes in either a DYT1 (KI) mice or a novel DYT1 transgenic model in open field, rotarod or staircase forepaw reaching tests. In the second part of this thesis we aimed to develop and evaluate the safety and efficacy of viral therapeutic RNAi constructs for in DYT1 murine models. DYT1 is an ideal candidate for this form of therapy due to its dominant inheritance, common mutation and potentially reversible phenotype. Virally delivered short-hairpin RNAs (shRNA) designed to knockdown torA(ÄE) in either an allele-specific or nonallele-specific manner were injected into the striatum of DYT1 transgenic or KI mice respectively. Unexpectedly, we found widespread lethal toxicity and behavioral abnormalities in mice injected with either therapeutic or control shRNAs that weren't observed in mice injected with no shRNAs. Further studies found that regions where toxic shRNAs were expressed corresponded with neuronal loss and glial activation. Finally, we found evidence that the severity of toxicity was influenced in part by the genetic background of the mice. In summary, the studies completed in this thesis contribute important information to the fields of dystonia… Advisors/Committee Members: Gonzalez-Alegre, Pedro (supervisor).

Subjects/Keywords: Dystonia; DYT1; RNAi; RNA interference; RNAi therapy; Genetics

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Martin, J. N. (2011). Developing RNAi therapy For DYT1 dystonia. (Doctoral Dissertation). University of Iowa. Retrieved from https://ir.uiowa.edu/etd/1019

Chicago Manual of Style (16^th Edition):

Martin, Janine Nicole. “Developing RNAi therapy For DYT1 dystonia.” 2011. Doctoral Dissertation, University of Iowa. Accessed January 18, 2021. https://ir.uiowa.edu/etd/1019.

MLA Handbook (7^th Edition):

Martin, Janine Nicole. “Developing RNAi therapy For DYT1 dystonia.” 2011. Web. 18 Jan 2021.

Vancouver:

Martin JN. Developing RNAi therapy For DYT1 dystonia. [Internet] [Doctoral dissertation]. University of Iowa; 2011. [cited 2021 Jan 18]. Available from: https://ir.uiowa.edu/etd/1019.

Council of Science Editors:

Martin JN. Developing RNAi therapy For DYT1 dystonia. [Doctoral Dissertation]. University of Iowa; 2011. Available from: https://ir.uiowa.edu/etd/1019

Penn State University

3. Kranick, Joshua Christian-issiah. Toward a More Complete Understanding of Microprocessing.

Degree: 2016, Penn State University

URL: https://submit-etda.libraries.psu.edu/catalog/27583

► Gene silencing via micro-RNAs is a cell’s main mechanism in maintaining cellular homeostasis. This is directly evidenced by the immense number of disease states directly… (more)

Subjects/Keywords: RNAi; miRNA; RNA; Microprocessing; Drosha; DGCR8; RNA Interference; miRNA Maturation

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Kranick, J. C. (2016). Toward a More Complete Understanding of Microprocessing. (Thesis). Penn State University. Retrieved from https://submit-etda.libraries.psu.edu/catalog/27583

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):

Kranick, Joshua Christian-issiah. “Toward a More Complete Understanding of Microprocessing.” 2016. Thesis, Penn State University. Accessed January 18, 2021. https://submit-etda.libraries.psu.edu/catalog/27583.

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

MLA Handbook (7^th Edition):

Kranick, Joshua Christian-issiah. “Toward a More Complete Understanding of Microprocessing.” 2016. Web. 18 Jan 2021.

Vancouver:

Kranick JC. Toward a More Complete Understanding of Microprocessing. [Internet] [Thesis]. Penn State University; 2016. [cited 2021 Jan 18]. Available from: https://submit-etda.libraries.psu.edu/catalog/27583.

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

Council of Science Editors:

Kranick JC. Toward a More Complete Understanding of Microprocessing. [Thesis]. Penn State University; 2016. Available from: https://submit-etda.libraries.psu.edu/catalog/27583

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

NSYSU

4. Wang, Ting-ya. Suppression of High Mobility Group Box-1 (HMGB-1) by RNAi Might Alter the Inflammatory Response During Sepsis.

Degree: Master, Biological Sciences, 2008, NSYSU

URL: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0904108-164845

► High mobility group box 1 (HMGB-1) protein is a non-histone chromosomal protein. As a DNA binding protein, HMGB-1 is involved in the maintenance of nucleosome… (more)

Subjects/Keywords: RNA interference (RNAi); sepsis; cytokine; High mobility group box 1 (HMGB1)

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Wang, T. (2008). Suppression of High Mobility Group Box-1 (HMGB-1) by RNAi Might Alter the Inflammatory Response During Sepsis. (Thesis). NSYSU. Retrieved from http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0904108-164845

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):

Wang, Ting-ya. “Suppression of High Mobility Group Box-1 (HMGB-1) by RNAi Might Alter the Inflammatory Response During Sepsis.” 2008. Thesis, NSYSU. Accessed January 18, 2021. http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0904108-164845.

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

MLA Handbook (7^th Edition):

Wang, Ting-ya. “Suppression of High Mobility Group Box-1 (HMGB-1) by RNAi Might Alter the Inflammatory Response During Sepsis.” 2008. Web. 18 Jan 2021.

Vancouver:

Wang T. Suppression of High Mobility Group Box-1 (HMGB-1) by RNAi Might Alter the Inflammatory Response During Sepsis. [Internet] [Thesis]. NSYSU; 2008. [cited 2021 Jan 18]. Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0904108-164845.

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

Council of Science Editors:

Wang T. Suppression of High Mobility Group Box-1 (HMGB-1) by RNAi Might Alter the Inflammatory Response During Sepsis. [Thesis]. NSYSU; 2008. Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0904108-164845

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

University of Melbourne

5. Pierce, Thomas Patrick. Use of RNAi in the mouse: inducible and reversible inhibition of Stat3.

Degree: 2012, University of Melbourne

URL: http://hdl.handle.net/11343/37193

► The signal transducer and activator of transcription 3 (Stat3) is a latent transcription factor that is activated during tissue inflammation and in a wide range… (more)

▼ The signal transducer and activator of transcription 3 (Stat3) is a latent transcription factor that is activated during tissue inflammation and in a wide range of human malignancies. Activation of Stat3 within the tumour microenvironment leads to the transcriptional induction of a network of Stat3 target genes. Among the transcriptional targets of Stat3 are genes that promote cell survival, cell cycle progression and factors that induce angiogenesis and suppress tumour surveillance by the adaptive immune system. Consequently, activation of Stat3 within the tumour microenvironment enables tumour cells to acquire several of the fundamental biological hallmarks of cancer. The Stat3 pathway therefore represents an attractive target for cancer therapeutics and inhibitors targeting several members of the pathway are currently under development. The work presented in this thesis aimed to develop pre-clinical mouse models in which the activity of Stat3 can be inhibited in a drug-like manner. A recently described method for RNA interference (RNAi)-based gene targeting was employed in order to achieve tetracycline (Tet)-inducible and reversible inhibition of Stat3 expression. To knockdown Stat3 expression, several short hairpin RNAs (shRNAs) were designed to specifically target the Stat3 mRNA. The efficacy of each of these shRNAs was assessed in two independent cell lines, using immunoblot analysis and Stat3 reporter gene assays to detect a reduction in Stat3 protein expression and function. The Stat3.1348 shRNA emerged as the most effective shRNA tested. Expression of Stat3.1348 in mouse embryonic stem cells resulted in cellular differentiation, providing additional evidence of effective Stat3 impairment by Stat3.1348. A mouse strain was generated in which a Tet-regulated Stat3.1348 construct was stably integrated into the genome at the collagen A1 locus. Furthermore, with the aim of facilitating tissue-specific Stat3 knockdown, a transgenic mouse strain harbouring an intestine-specific Tet-transactivator was generated and the function of this tissue-specific Tet-transactivator was compared to two published Tet-transactivator strains with more ubiquitous expression patterns. Mice harbouring the Stat3.1348 allele were crossed with Tet-transactivator strain with the most rebust intestinal activity, R26-M2rtTA. No reduction in Stat3 protein expression or function was detected in the gastrointestinal tract after treatment of R26-M2rtTA/Stat3.1348 mice with doxycycline to induce shRNA expression. In a parallel study, the utility of a retrovirally-delivered Stat3.1348 construct targeting Stat3 was explored in the mouse 4T1 breast tumour model. This tumour cell line exhibits constitutive Stat3 activity and 4T1 tumour growth in the mouse mammary fat pad has been previously been shown to dependent on Stat3 expression. However, despite achieving effective reduction of Stat3 expression in 4T1 cells using retroviral Stat3.1348 expression constructs, tumour growth was not found to be altered…

Subjects/Keywords: RNAi; RNA interference; shRNA; Stat3; miR-21; mouse models; gene knockdown

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Pierce, T. P. (2012). Use of RNAi in the mouse: inducible and reversible inhibition of Stat3. (Doctoral Dissertation). University of Melbourne. Retrieved from http://hdl.handle.net/11343/37193

Chicago Manual of Style (16^th Edition):

Pierce, Thomas Patrick. “Use of RNAi in the mouse: inducible and reversible inhibition of Stat3.” 2012. Doctoral Dissertation, University of Melbourne. Accessed January 18, 2021. http://hdl.handle.net/11343/37193.

MLA Handbook (7^th Edition):

Pierce, Thomas Patrick. “Use of RNAi in the mouse: inducible and reversible inhibition of Stat3.” 2012. Web. 18 Jan 2021.

Vancouver:

Pierce TP. Use of RNAi in the mouse: inducible and reversible inhibition of Stat3. [Internet] [Doctoral dissertation]. University of Melbourne; 2012. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/11343/37193.

Council of Science Editors:

Pierce TP. Use of RNAi in the mouse: inducible and reversible inhibition of Stat3. [Doctoral Dissertation]. University of Melbourne; 2012. Available from: http://hdl.handle.net/11343/37193

University of Edinburgh

6. Castonguay, Emilie. Identification of novel inhibitors of heterochromatin integrity through a chemical screen in fission yeast.

Degree: PhD, 2014, University of Edinburgh

URL: http://hdl.handle.net/1842/17563

► Heterochromatin assembly in fission yeast (Schizosaccharomyces pombe) requires conserved components that mediate RNA interference (RNAi) directed methylation of histone H3 on lysine 9 (H3K9). Fission… (more)

▼ Heterochromatin assembly in fission yeast (Schizosaccharomyces pombe) requires conserved components that mediate RNA interference (RNAi) directed methylation of histone H3 on lysine 9 (H3K9). Fission yeast heterochromatin is mainly found at centromeres, telomeres, and the mating-type locus. At centromeres, transcripts from repetitive elements are processed to siRNAs and RNAi promotes chromatin modification by recruiting the Clr4 methyltransferase. RNAi is not required to maintain silent chromatin at the mating-type locus. This RNAi-directed form of centromeric heterochromatin provides an ideal system for in vivo screening to allow the identification of compounds that inhibit the activity of proteins involved in RNA silencing, chromatin modification and heterochromatin assembly in fission yeast and may inhibit conserved proteins in other organisms. A dominant selectable marker gene system at fission yeast centromeres that reports loss of heterochromatin integrity by increased resistance to G418 in 96-well plate format liquid cultures was developed. The resulting strain was used to screen a nontargeted chemically diverse compound library in vivo to identify compounds that disrupt the integrity of RNAi-directed heterochromatin. Two compounds, Emi1 and Emi14, were identified and found to cause a significant decrease in the level of H3K9 methylation on the outer repeats at fission yeast centromeres. Growth in the presence of Emi1 or Emi14 also caused a reduction in H3K9 methylation levels at the mating-type locus, suggesting that they do not act through RNAi. Consistent with this, Emi1 and Emi14 did not cause a decrease in centromeric siRNA levels. Analyses therefore suggest that Emi1 and Emi14 do not disrupt RNAi but that they inhibit downstream events in chromatin modification and heterochromatin assembly. Cells lacking RNAi due to loss of Dicer (dcr1Δ) or cells lacking the histone deacetylase (HDAC) Sir2 (sir2Δ) retain significant but lower levels of H3K9 methylation on the centromeric outer repeats. When dcr1Δ or sir2Δ cells were grown in the presence of Emi1 or Emi14 a further reduction in H3K9 methylation levels was observed on the outer repeats. This mimics the effect of combining clr3Δ with dcr1Δ or sir2Δ and suggests that Emi1 and Emi14 may interfere with SHREC function. SHREC is a chromatin remodelling complex that includes the HDAC Clr3 and the chromatin remodeler Mit1 and is known to contribute to heterochromatin integrity. Expression profiling performed on Emi1 and Emi14 treated cells confirmed the previous results. The changes in gene expression following Emi1 and Emi14 treatment were compared to known mutants defective in heterochromatin integrity. The profile of expression changes following Emi14 treatment was found to correlate with alterations in the expression pattern observed in cells with SHREC components deleted. No correlation with mutants lacking other HDACs or RNAi components was detected. Emi1 had a weaker correlation with defective SHREC function and thus may also partially inhibit the SHREC…

Subjects/Keywords: 572; heterochromatin; RNA interference; RNAi; chemical biology; fission yeast; chromatin

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Castonguay, E. (2014). Identification of novel inhibitors of heterochromatin integrity through a chemical screen in fission yeast. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/17563

Chicago Manual of Style (16^th Edition):

Castonguay, Emilie. “Identification of novel inhibitors of heterochromatin integrity through a chemical screen in fission yeast.” 2014. Doctoral Dissertation, University of Edinburgh. Accessed January 18, 2021. http://hdl.handle.net/1842/17563.

MLA Handbook (7^th Edition):

Castonguay, Emilie. “Identification of novel inhibitors of heterochromatin integrity through a chemical screen in fission yeast.” 2014. Web. 18 Jan 2021.

Vancouver:

Castonguay E. Identification of novel inhibitors of heterochromatin integrity through a chemical screen in fission yeast. [Internet] [Doctoral dissertation]. University of Edinburgh; 2014. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/1842/17563.

Council of Science Editors:

Castonguay E. Identification of novel inhibitors of heterochromatin integrity through a chemical screen in fission yeast. [Doctoral Dissertation]. University of Edinburgh; 2014. Available from: http://hdl.handle.net/1842/17563

7. Grypioti, Emilia. Identification and functional characterization of RNA silencing key-genes in the model pennate diatom species Phaeodactylum tricornutum.

Degree: 2019, University of Crete (UOC); Πανεπιστήμιο Κρήτης

URL: http://hdl.handle.net/10442/hedi/46845

► Gene silencing, also known as RNA interference (RNAi), is a conserved mechanism of regulation of gene expression mediated by small RNAs (sRNA), (Fire et al.,… (more)

▼ Gene silencing, also known as RNA interference (RNAi), is a conserved mechanism of regulation of gene expression mediated by small RNAs (sRNA), (Fire et al., 1998). Silencing of transgenes and endogenous genes following introduction of inverted repeats, antisense constructs and artificial miRNAs has been reported in diatom species, including the model species P. tricornutum (De Riso et al., 2009; Kaur and Spillane, 2015). The presence of an endogenous RNAi pathway has been suggested after comprehensive and combinatorial analyses of sRNAs, gene expression and DNA methylation in P. tricornutum (Veluchamy et al., 2013; Rogato et al., 2014). This RNAi pathway in P. tricornutum may play a role in the regulation of protein coding genes and TEs expression with possible consequences for the acclamatory response to nutrient limitation (Maumus et al., 2009). Homologues of the RNAi-key genes DICER (DCR), ARGONAUTE (AGO) and RNA-Dependent RNA polymerase (RDR) have been previously identified by in silico analysis (De Riso et al., 2009). However, the validation of their gene models, the characterization of their functions and the possible physiological role of RNAi in diatoms are still lacking. In this study, extensive in silico analysis of genomic and transcriptomic information available in P. tricornutum suggests the presence of a single PtDCR, PtAGO and PtRDR coding gene. Mining and phylogenetic analysis of DCR, AGO and RDR homologues in diatoms from all publically available to date sequence datasets suggest an unanticipated diversification of the RNAi pathway in these organisms. PtDCR/AGO/RDR cDNA were cloned and splicing isoforms of PtDCR and PtAGO were identified. Subcellular localization of PtDCR-/AGO-/RDR-YFP was investigated by confocal microscopy. Functional characterization of PtDCR and PtAGO was first attempted by heterologous expression in the yeast Saccharomyces cerevisiae and the plant Nicotiana bethamiana hosts. In a second step, CRISPR/Cas9-mediated mutagenesis approach, recently developed in P. tricornutum, was successfully harnessed to generate PtDCR-KO and PtAGO-KO (KnockOut) lines. Growth phenotype of PtDCR-KO lines were investigated under optimal and nitrate depleted culture conditions and during recovery from UV-mediated stress. In parallel, mRNA and small RNAs whole transcriptome analyses were carried out. Culture experiments suggest that PtDCR may play a role in the response to nitrate starvation. Transcriptomic analysis revealed that both sRNA and mRNA transcriptomes were affected in PtDCR-KO line. At the global scale, sRNA size distribution was found to shift towards larger fragment size in PtDCR-KO line. In addition, the abundance of sRNA mapped to TEs was found dramatically reduced in PtDCR-KO mutant and a concomitant increase in mRNA abundance of some TEs was observed. Interestingly, PtDCR-KO sRNA transcriptome also presented changes in tRNA-derived sRNA populations, suggesting a possible role of DCR in their processing in diatoms. TE mobilization has been proposed to play a pivotal role in diatom…

Subjects/Keywords: Διάτομα; Ρύθμιση γονιδιακής έκφρασης; Diatoms; RNA interference (RNAi); Phaeodactylum tricornutum

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Grypioti, E. (2019). Identification and functional characterization of RNA silencing key-genes in the model pennate diatom species Phaeodactylum tricornutum. (Thesis). University of Crete (UOC); Πανεπιστήμιο Κρήτης. Retrieved from http://hdl.handle.net/10442/hedi/46845

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):

Grypioti, Emilia. “Identification and functional characterization of RNA silencing key-genes in the model pennate diatom species Phaeodactylum tricornutum.” 2019. Thesis, University of Crete (UOC); Πανεπιστήμιο Κρήτης. Accessed January 18, 2021. http://hdl.handle.net/10442/hedi/46845.

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

MLA Handbook (7^th Edition):

Grypioti, Emilia. “Identification and functional characterization of RNA silencing key-genes in the model pennate diatom species Phaeodactylum tricornutum.” 2019. Web. 18 Jan 2021.

Vancouver:

Grypioti E. Identification and functional characterization of RNA silencing key-genes in the model pennate diatom species Phaeodactylum tricornutum. [Internet] [Thesis]. University of Crete (UOC); Πανεπιστήμιο Κρήτης; 2019. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/10442/hedi/46845.

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

Council of Science Editors:

Grypioti E. Identification and functional characterization of RNA silencing key-genes in the model pennate diatom species Phaeodactylum tricornutum. [Thesis]. University of Crete (UOC); Πανεπιστήμιο Κρήτης; 2019. Available from: http://hdl.handle.net/10442/hedi/46845

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

Kansas State University

8. Cooper, Anastasia M. W. Molecular mechanisms influencing the efficiency of RNA interference in the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae).

Degree: PhD, Department of Entomology, 2020, Kansas State University

URL: http://hdl.handle.net/2097/40905

► RNA interference (RNAi) pathways function in endogenous gene regulation and protection against viruses and transposons in eukaryotic organisms. The exogenous RNAi mechanism has been utilized… (more)

▼ RNA interference (RNAi) pathways function in endogenous gene regulation and protection against viruses and transposons in eukaryotic organisms. The exogenous RNAi mechanism has been utilized as a powerful reverse genetics tool to analyze gene function. In addition, RNAi-based pest management strategies are now emerging; however, applications are limited due to inefficient results for some insects, especially lepidopterans. The European corn borer (ECB), Ostrinia nubilalis, is an agriculturally relevant lepidopteran pest that exhibits very low RNAi efficiency but has a great need for new control strategies that utilize novel modes of action. This dissertation research seeks to investigate molecular mechanisms influencing RNAi efficiency in ECB and determine if commonly used reagents and tactics can improve RNAi efficiency in this species. The specific objectives were: 1) to investigate double-stranded RNA (dsRNA) stability in larval gut contents and hemolymph in ECB, 2) to identify and characterize dsRNA-degrading nuclease genes from ECB, 3) to identify and characterize core RNAi pathway genes from ECB, and 4) to compare strategies for enhancing RNAi efficiency (nanoparticles, transfection reagents, and nuclease inhibitors) in ECB. Ex vivo incubation experiments showed that dsRNA was degraded in ECB gut contents and hemolymph under physiologically relevant pH conditions. Sequencing revealed transcripts for four dsRNA-degrading endonuclease (OndsRNase) genes and one RNAi efficiency-related nuclease (OnREase) gene in ECB. Expression analysis indicated that OnREase, OndsRNase2, and OndsRNase4 might impact oral RNAi efficiency in ECB, whereas OnREase and OndsRNase1 may impact injection RNAi efficiency. Sequencing also revealed a single transcript for Dicer 2 (OnDcr2), R2D2 (OnR2D2), and Argonaute 2 (OnAgo2). Expression analysis indicated that each of these core RNAi pathway genes were expressed in all ECB developmental stages and tissues investigated. Ex vivo incubation experiments showed that Metafectene Pro, chitosan-based nanoparticles, EDTA, and Zn2+ enhanced dsRNA stability in ECB gut contents and hemolymph, but Lipofectamine RNAiMax, Co2+, and Mn2+ did not. Surprisingly, improving dsRNA stability did not increase RNAi efficiency in ECB in vivo. Ingestion, injection, and tissue culture-based RNAi assays indicated that most target genes and all ECB developmental stages investigated were highly refractory to RNAi. These findings suggest that multiple mechanisms, including instability of dsRNA, deficient core RNAi machinery, and refractory target genes, likely contribute to low RNAi efficiency in ECB. Primary outcomes of this project included an in-depth investigation of dsRNA instability under physiological conditions in ECB tissues, a preliminary identification of candidate dsRNA-degrading nuclease genes that likely contribute to dsRNA instability in ECB, a foundational characterization of core RNAi pathway genes presumed to participate in the exogenous RNAi response of ECB, and a detailed comparison of numerous… Advisors/Committee Members: Kun Yan Zhu.

Subjects/Keywords: RNA interference; Insect bioassays; Double-stranded RNA stability; Nucleases; Core RNAi pathway genes; Target genes

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Cooper, A. M. W. (2020). Molecular mechanisms influencing the efficiency of RNA interference in the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae). (Doctoral Dissertation). Kansas State University. Retrieved from http://hdl.handle.net/2097/40905

Chicago Manual of Style (16^th Edition):

Cooper, Anastasia M W. “Molecular mechanisms influencing the efficiency of RNA interference in the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae).” 2020. Doctoral Dissertation, Kansas State University. Accessed January 18, 2021. http://hdl.handle.net/2097/40905.

MLA Handbook (7^th Edition):

Cooper, Anastasia M W. “Molecular mechanisms influencing the efficiency of RNA interference in the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae).” 2020. Web. 18 Jan 2021.

Vancouver:

Cooper AMW. Molecular mechanisms influencing the efficiency of RNA interference in the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae). [Internet] [Doctoral dissertation]. Kansas State University; 2020. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/2097/40905.

Council of Science Editors:

Cooper AMW. Molecular mechanisms influencing the efficiency of RNA interference in the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae). [Doctoral Dissertation]. Kansas State University; 2020. Available from: http://hdl.handle.net/2097/40905

9. Kelli Cristina Micocci. Análise da expressão e do papel da ADAM9 humana na capacidade invasiva de células tumorais por silenciamento gênico.

Degree: 2009, Universidade Federal de São Carlos

URL: http://www.bdtd.ufscar.br/htdocs/tedeSimplificado//tde_busca/arquivo.php?codArquivo=3267

►

ADAMs é um termo usado para descrever a presença de domínios desintegrina emetaloprotease (A Disintegrin And Metalloprotease) em uma determinada classe de proteínasde membrana, multifuncionais,… (more)

▼

ADAMs é um termo usado para descrever a presença de domínios desintegrina emetaloprotease (A Disintegrin And Metalloprotease) em uma determinada classe de proteínasde membrana, multifuncionais, expressas em diferentes espécies animais como mamíferos einsetos. Elas possuem funções importantes em muitos processos fisiológicos como nafertilização, fusão de mioblastos, migração, proliferação e sobrevivência celular, entre outros,bem como em processos patológicos tais como muitos tipos de tumores humanos, incluindomama, próstata, pâncreas, fígado, rins e pele. A ADAM9 está envolvida em diversosprocessos celulares, tais como adesão celular, migração e sinalização de células tumorais,contribuindo para o desenvolvimento de metástases. Objetivos: Analisar a expressão daADAM9 em linhagens de células tumorais (MDA-MB-231 e DU-145) e não tumorais (FH eC2C12), gerar clones com o gene que codifica para a ADAM9 silenciados e verificar o efeitoda ausência desta proteína na invasão, proliferação e expressão gênica das células MDA-MB-231. Métodos: As células foram cultivadas e posteriormente plaqueadas (2x106 células/placade 6cm) por 24 horas e em 5ml de meio de cultura DMEM (10% de FBS) e lisadas para oensaio de western blotting e zimografia. Para o ensaio de inibição de adesão as células MDAMB-231 e FH (5x106 células/ml) foram marcadas com CMFDA (clorometil diacetatofluoresceína) e posteriormente incubadas com os anticorpos anti-ADAM9D e anti-RP3ADAM9 em diferentes concentrações. Para a técnica de RNAi foi utilizado um kit(Silencer siRNA Starter Kit Ambion), 2,0x105 de células (MDA-MB-231) e agente detransfecção (Lipofectamina 2000 - Invitrogen). As células foram plaqueadas em 5ml de meiode cultura DMEM/placa. No terceiro dia de plaqueamento as células foram tratadas comagente de transfecção e primer de silenciamento do RNA, para serem utilizadas na RT-PCR,ensaio de proliferação e invasão em matrigel. Resultados: Todas as linhagens estudadasexpressaram a ADAM9 nas condições de estudo. A atividade MMP-2 (gelatinase-A)intermediária estava presente em todos os tipos celulares testados. O silenciamento deADAM9 não afetou a taxa de proliferação das células MDA-MB-231 após 4, 5, 6, 7, 8, 9 e 10dias do silenciamento (24 ou 48 horas de incubação). O silenciamento da ADAM9 humanainibiu a invasão celular em células de câncer de mama (MDA-MB-231) em matrigel (71,51 8,02%) quando comparados com o controle. Conclusão: A geração de clones knockout sem aexpressão da ADAM9 utilizando a técnica de silenciamento de RNA em células de câncer demama (MDA-MB-231), não afetou a taxa de proliferação celular. No entanto, a invasão dascélulas tumorais em matrigel foi inibida em aproximadamente 70% quando comparada com ocontrole, demonstrando que ADAM9 é uma importante molécula envolvida no processo deinvasão e metástase.

ADAMs is a term used to describe the presence of disintegrin and metalloprotease domains(A Disintegrin And Metalloprotease) in a certain class of multi-functional membrane proteins,expressed in several animal species such as mammals and…

Advisors/Committee Members: Heloísa Sobreiro Selistre de Araujo.

Subjects/Keywords: Câncer e RNA de interferência (RNAi); ADAM9; Fisiologia; RNA interferente; Mamas - câncer; Bioquímica; FISIOLOGIA; ADAM9; Cancer and interference RNA (iRNA)

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Micocci, K. C. (2009). Análise da expressão e do papel da ADAM9 humana na capacidade invasiva de células tumorais por silenciamento gênico. (Thesis). Universidade Federal de São Carlos. Retrieved from http://www.bdtd.ufscar.br/htdocs/tedeSimplificado//tde_busca/arquivo.php?codArquivo=3267

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):

Micocci, Kelli Cristina. “Análise da expressão e do papel da ADAM9 humana na capacidade invasiva de células tumorais por silenciamento gênico.” 2009. Thesis, Universidade Federal de São Carlos. Accessed January 18, 2021. http://www.bdtd.ufscar.br/htdocs/tedeSimplificado//tde_busca/arquivo.php?codArquivo=3267.

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

MLA Handbook (7^th Edition):

Micocci, Kelli Cristina. “Análise da expressão e do papel da ADAM9 humana na capacidade invasiva de células tumorais por silenciamento gênico.” 2009. Web. 18 Jan 2021.

Vancouver:

Micocci KC. Análise da expressão e do papel da ADAM9 humana na capacidade invasiva de células tumorais por silenciamento gênico. [Internet] [Thesis]. Universidade Federal de São Carlos; 2009. [cited 2021 Jan 18]. Available from: http://www.bdtd.ufscar.br/htdocs/tedeSimplificado//tde_busca/arquivo.php?codArquivo=3267.

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

Council of Science Editors:

Micocci KC. Análise da expressão e do papel da ADAM9 humana na capacidade invasiva de células tumorais por silenciamento gênico. [Thesis]. Universidade Federal de São Carlos; 2009. Available from: http://www.bdtd.ufscar.br/htdocs/tedeSimplificado//tde_busca/arquivo.php?codArquivo=3267

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

University of Florida

10. Fitzpatrick, Daniel M. Escape from Short-Interfering RNA-Induced Silencing in an Orthobunyavirus, Tensaw Virus.

Degree: MS, Entomology and Nematology, 2013, University of Florida

URL: https://ufdc.ufl.edu/UFE0045564

► Ribonucleic acid interference (RNAi) is a major component of antiviral immunity in dipteran insects, including mosquitoes. Virus-specific short-interfering RNA (siRNA) are being considered as a… (more)

Subjects/Keywords: Antivirals; Cell growth; Hela cells; Infections; Nonsense; RNA; RNA interference; Small interfering RNA; Vero cells; Viruses; arbovirus – entomology – mosquito – rnai – virus

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Fitzpatrick, D. M. (2013). Escape from Short-Interfering RNA-Induced Silencing in an Orthobunyavirus, Tensaw Virus. (Masters Thesis). University of Florida. Retrieved from https://ufdc.ufl.edu/UFE0045564

Chicago Manual of Style (16^th Edition):

Fitzpatrick, Daniel M. “Escape from Short-Interfering RNA-Induced Silencing in an Orthobunyavirus, Tensaw Virus.” 2013. Masters Thesis, University of Florida. Accessed January 18, 2021. https://ufdc.ufl.edu/UFE0045564.

MLA Handbook (7^th Edition):

Fitzpatrick, Daniel M. “Escape from Short-Interfering RNA-Induced Silencing in an Orthobunyavirus, Tensaw Virus.” 2013. Web. 18 Jan 2021.

Vancouver:

Fitzpatrick DM. Escape from Short-Interfering RNA-Induced Silencing in an Orthobunyavirus, Tensaw Virus. [Internet] [Masters thesis]. University of Florida; 2013. [cited 2021 Jan 18]. Available from: https://ufdc.ufl.edu/UFE0045564.

Council of Science Editors:

Fitzpatrick DM. Escape from Short-Interfering RNA-Induced Silencing in an Orthobunyavirus, Tensaw Virus. [Masters Thesis]. University of Florida; 2013. Available from: https://ufdc.ufl.edu/UFE0045564

Queensland University of Technology

11. Mware, Benard Ouma. Development of banana bunchy top virus resistance in bananas: RNAi approach.

Degree: 2016, Queensland University of Technology

URL: http://eprints.qut.edu.au/95736/

► Bunchy top, caused by banana bunchy top virus (BBTV), is the most important viral disease affecting banana worldwide, for which control is very difficult. In… (more)

Subjects/Keywords: Banana; Banana bunchy top virus; RNA interference; Hairpin; Transgenic plants; siRNA; Musa; RNAi

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Mware, B. O. (2016). Development of banana bunchy top virus resistance in bananas: RNAi approach. (Thesis). Queensland University of Technology. Retrieved from http://eprints.qut.edu.au/95736/

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):

Mware, Benard Ouma. “Development of banana bunchy top virus resistance in bananas: RNAi approach.” 2016. Thesis, Queensland University of Technology. Accessed January 18, 2021. http://eprints.qut.edu.au/95736/.

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

MLA Handbook (7^th Edition):

Mware, Benard Ouma. “Development of banana bunchy top virus resistance in bananas: RNAi approach.” 2016. Web. 18 Jan 2021.

Vancouver:

Mware BO. Development of banana bunchy top virus resistance in bananas: RNAi approach. [Internet] [Thesis]. Queensland University of Technology; 2016. [cited 2021 Jan 18]. Available from: http://eprints.qut.edu.au/95736/.

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

Council of Science Editors:

Mware BO. Development of banana bunchy top virus resistance in bananas: RNAi approach. [Thesis]. Queensland University of Technology; 2016. Available from: http://eprints.qut.edu.au/95736/

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

Universiteit Utrecht

12. Steiner, F.A. Small RNAs and Argonaute proteins: key players in post-transcriptional gene silencing.

Degree: 2007, Universiteit Utrecht

URL: http://dspace.library.uu.nl:8080/handle/1874/23784

► Small RNAs are important transcriptional and post-transcriptional regulators of gene expression. Many classes of small RNAs have been discovered, each carrying out specialized functions. siRNAs… (more)

▼ Small RNAs are important transcriptional and post-transcriptional regulators of gene expression. Many classes of small RNAs have been discovered, each carrying out specialized functions. siRNAs and miRNAs are best studies. siRNAs function in the process of RNAi and are thought to defend the genome against molecular parasites. They have become widely used tools in biomedical research. miRNAs are abundant and conserved; they form a class of endogenous small RNAs that fine-tunes gene expression. Most, if not all, small RNAs function through association with members of the Argonaute protein family. This thesis focuses on the function of small RNAs and Argonaute proteins in RNAi and the miRNA pathway. To understand the role of miRNAs in development, we aimed to catalogue the zebrafish miRNAs and analyze their expression. By sequencing miRNA libraries from 5-day-old zebrafish larvae and adult zebrafish brain, we found 139 known and 66 new miRNAs. We analyzed the temporal and spatial expression patterns of 67 miRNAs by whole mount in situ hybridization and northern blot analysis. Most miRNAs are expressed during later stages of development, often in a tissue specific manner. Most newly discovered miRNAs have low expression levels and are less conserved in other vertebrate species. We also analyzed the connection between the miRNAs and Argonaute proteins in C. elegans. The Argonaute proteins required for the miRNA pathway are ALG-1 and ALG-2. We used massively parallel sequencing to analyze the spectrum of small RNAs associated with ALG-1. We found that this protein almost exclusively binds to miRNAs. In addition to 84 known miRNAs, we found ten novel miRNAs. RNAi is an amplified process in C. elegans. In the amplification step, RNA-directed RNA polymerases (RdRPs) produce secondary siRNAs, which we cloned from transgenic lines expressing a single primary siRNA. We found that RdRPs perform unprimed RNA synthesis to generate secondary siRNAs, and that mRNAs which are not cleaved by the primary siRNA-Argonaute complex can be used as substrates. Secondary siRNAs are only of antisense polarity, carry 5' di- or triphosphates, and are only in the minority associated with RDE-1, the primary siRNA-binding Argonaute protein. Therefore, secondary siRNAs represent a distinct class of small RNAs. siRNAs and miRNAs are both processed from dsRNA-precursors by the dsRNase DCR-1. We investigated how RNAi and the miRNA pathway are mechanistically separated. We showed that precursors of small RNAs contain structural features that direct the small RNAs into the RNAi or the miRNA pathway. Small RNAs expressed from hairpin precursors with a fully matching stem are recognized as siRNAs and bound by RDE-1. A one- to three-nucleotide mismatch at various positions in the stem of the precursor directs the small RNAs into the miRNA pathway, as these small RNAs are in majority bound to ALG-1. The Argonaute proteins to which the small RNAs are bound determine the silencing mode, and no functional overlap between RDE-1 and ALG-1 was detected. We also…

Subjects/Keywords: Biologie; RNA interference; RNAi; miRNA; Argonaute; RdRP; Caenorhabditis elegans; RDE-1; ALG-1; zebrafish

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Steiner, F. A. (2007). Small RNAs and Argonaute proteins: key players in post-transcriptional gene silencing. (Doctoral Dissertation). Universiteit Utrecht. Retrieved from http://dspace.library.uu.nl:8080/handle/1874/23784

Chicago Manual of Style (16^th Edition):

Steiner, F A. “Small RNAs and Argonaute proteins: key players in post-transcriptional gene silencing.” 2007. Doctoral Dissertation, Universiteit Utrecht. Accessed January 18, 2021. http://dspace.library.uu.nl:8080/handle/1874/23784.

MLA Handbook (7^th Edition):

Steiner, F A. “Small RNAs and Argonaute proteins: key players in post-transcriptional gene silencing.” 2007. Web. 18 Jan 2021.

Vancouver:

Steiner FA. Small RNAs and Argonaute proteins: key players in post-transcriptional gene silencing. [Internet] [Doctoral dissertation]. Universiteit Utrecht; 2007. [cited 2021 Jan 18]. Available from: http://dspace.library.uu.nl:8080/handle/1874/23784.

Council of Science Editors:

Steiner FA. Small RNAs and Argonaute proteins: key players in post-transcriptional gene silencing. [Doctoral Dissertation]. Universiteit Utrecht; 2007. Available from: http://dspace.library.uu.nl:8080/handle/1874/23784

Stellenbosch University

13. Visser, Ilze. RNAi of selected insect genes.

Degree: MSc, Genetics, 2016, Stellenbosch University

URL: http://hdl.handle.net/10019.1/100274

► ENGLISH ABSTRACT: Diuraphis noxia (Kurdjumov, Hemiptera: Aphididae), commonly known as the Russian wheat aphid (RWA), is regarded as one of the most destructive and widely… (more)

▼ ENGLISH ABSTRACT: Diuraphis noxia (Kurdjumov, Hemiptera: Aphididae), commonly known as the Russian wheat aphid (RWA), is regarded as one of the most destructive and widely distributed insect species in the world. Nonetheless, the currently available control strategies, including chemical pesticides, biological control agents, and RWA resistant wheat cultivars, are still very limited and rather ineffective. The process of double-stranded RNA (dsRNA)-mediated interference (RNAi) displays high specificity and the prospect of developing into a new specific method for managing agricultural pests. Plants can potentially be genetically engineered to express dsRNA to down-regulate vital gene functions present in pest insects, resulting in the protection of plants. In order to survive and reproduce, aphids require close interaction with their host plants, during which effectors are transported inside the plant to modify host cell processes. Four previously identified RWA salivary secretion proteins were investigated in the present study. However, cloning and sequencing results indicated that only two of the aforementioned proteins – C002 and 14-3-3 epsilon (ɛ) – could be potential protein elicitors in RWA. Thus, these two transcripts were subjected to RNAi experiments via artificial diet feeding and feeding on siRNA injected wheat leaf trials in order to investigate their role in RWA-host interactions and their importance in the survival and reproduction of the RWA. The relative expression levels of C002 and 14-3-3 ɛ at 0h were compared between SAM, the most virulent RWA biotype, and SA1, the least virulent RWA biotype in South Africa, and the results indicated that both transcripts had a higher relative expression in SAM than in SA1. Therefore, suggesting that C002 and 14-3-3 ɛ might play an important part in RWA virulence. From the RT-qPCR results it was evident that successful silencing of both C002 and 14-3-3 ɛ were achieved at 24h after initial siRNA exposure and that the transient silencing effect subsided thereafter. The expression data pertaining to the wheat leaf injection experiments, however, displayed high standard deviations that are not ideal and suggested that the expression of the transcripts differs greatly between the aphids within each group. This is likely due to the custom-made aphid cages and injection procedure of the siRNA into wheat leaves that appears to hinder the accuracy of the results. The fecundity data produced quite inconclusive results due to previously mentioned inadequacies and therefore an accurate and decisive conclusion cannot be drawn as to how the C002 and 14-3-3 ɛ silencing effects the survival and reproduction of the RWA. Both methods used for RNAi – the artificial diet trial and the injection of wheat leaves trial – have their drawback. After considering the RT-qPCR data, it appears as though the artificial diet trial produced more accurate and feasible results. Even so, the injection method establishes a more natural mode of feeding for the aphids and consequently more optimal… Advisors/Committee Members: Botha-Oberholster, A-M., Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics..

Subjects/Keywords: Russian wheat aphid (RWA); Double-stranded RNA (dsRNA)-mediated interference (RNAi); Insect pest control; UCTD

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Visser, I. (2016). RNAi of selected insect genes. (Masters Thesis). Stellenbosch University. Retrieved from http://hdl.handle.net/10019.1/100274

Chicago Manual of Style (16^th Edition):

Visser, Ilze. “RNAi of selected insect genes.” 2016. Masters Thesis, Stellenbosch University. Accessed January 18, 2021. http://hdl.handle.net/10019.1/100274.

MLA Handbook (7^th Edition):

Visser, Ilze. “RNAi of selected insect genes.” 2016. Web. 18 Jan 2021.

Vancouver:

Visser I. RNAi of selected insect genes. [Internet] [Masters thesis]. Stellenbosch University; 2016. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/10019.1/100274.

Council of Science Editors:

Visser I. RNAi of selected insect genes. [Masters Thesis]. Stellenbosch University; 2016. Available from: http://hdl.handle.net/10019.1/100274

University of Edinburgh

14. Chapman, Elliott. Investigating the role of RNA interference in the fission yeast Schizosaccharomyces japonicus.

Degree: PhD, 2018, University of Edinburgh

URL: http://hdl.handle.net/1842/31201

► RNA interference (RNAi) is a conserved pathway that plays key roles in heterochromatin formation, gene regulation and genome surveillance across a wide range of eukaryotes.… (more)

▼ RNA interference (RNAi) is a conserved pathway that plays key roles in heterochromatin formation, gene regulation and genome surveillance across a wide range of eukaryotes. One of the most utilised model organisms for studying the RNAi pathway is the fission yeast Schizosaccharomyces pombe. However, this species is somewhat atypical, in that it has not retained the ancestral role for RNAi in the silencing of mobile genetic elements. In contrast, the related fission yeast S. japonicus has a large and diverse retrotransposon complement that appears to give rise to abundant siRNAs. For this reason, we believe that S. japonicus may be a more suitable model for studying the role of RNAi in silencing mobile genetic elements, a function that is conserved in many higher eukaryotes. Functional analysis of the S. japonicus RNAi pathway proved more challenging than expected, as it was generally not possible to recover strains bearing deletions of core RNAi components (Ago1/Clr4/Rdp1/Arb1/Arb2). This suggests that a functional RNAi pathway may be required for viability in S. japonicus, unlike in S. pombe. However, disruption mutants were isolated for the sole Dicer ribonuclease Dcr1, at very low frequency. Analysis of these mutants revealed that disruption of Dcr1 impaired the generation of retrotransposon derived siRNAs, and caused de-repression of retroelement transcript accumulation and mobilisation in an element dependent manner. Surprisingly however, Dcr1 appeared dispensable for the maintenance of H3K9me2 at transposons, suggesting that, in contrast to S. pombe, silencing may occur principally at the post-transcriptional level. It is also possible that the isolated Dcr1 mutants represent rare survivors that are viable due to the presence of suppressor mutations elsewhere in the genome. I utilised my genome wide RNA sequencing data to help improve the annotation of the S. japonicus genome, with a specific focus on the retrotransposon complement. From this, I identified 12 new families of LTR retrotransposon, which increased the annotated retrotransposon complement by around 40% in S. japonicus. Finally, I characterised the integrative preference of the S. japonicus retrotransposon Tj1, and found that it shares characteristics associated with the S. cerevisiae retrotransposons Ty1 and Ty3, mostly integrating upstream of RNA PolIII transcribed tRNA genes. The findings of this work highlight some potentially key differences in the way the RNAi pathway functions across the fission yeast clade, both in terms of its importance for viability and its mode of action. The work undertaken here also contributes to the establishment of S. japonicus as a model for the study of RNA interference and genome regulation.

Subjects/Keywords: 572.8; chromatin; heterochromatin; RNA interference; RNAi; S. japonicus; Schizosaccharomyces pombe; Schizosaccharomyces japonicus; post-transcription

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Chapman, E. (2018). Investigating the role of RNA interference in the fission yeast Schizosaccharomyces japonicus. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/31201

Chicago Manual of Style (16^th Edition):

Chapman, Elliott. “Investigating the role of RNA interference in the fission yeast Schizosaccharomyces japonicus.” 2018. Doctoral Dissertation, University of Edinburgh. Accessed January 18, 2021. http://hdl.handle.net/1842/31201.

MLA Handbook (7^th Edition):

Chapman, Elliott. “Investigating the role of RNA interference in the fission yeast Schizosaccharomyces japonicus.” 2018. Web. 18 Jan 2021.

Vancouver:

Chapman E. Investigating the role of RNA interference in the fission yeast Schizosaccharomyces japonicus. [Internet] [Doctoral dissertation]. University of Edinburgh; 2018. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/1842/31201.

Council of Science Editors:

Chapman E. Investigating the role of RNA interference in the fission yeast Schizosaccharomyces japonicus. [Doctoral Dissertation]. University of Edinburgh; 2018. Available from: http://hdl.handle.net/1842/31201

University of Georgia

15. Andersen, Lauren Elizabeth. MiRNA regulation and human protein kinase genes required for influenza virus replication.

Degree: 2014, University of Georgia

URL: http://hdl.handle.net/10724/27660

► Human protein kinases (HPKs) have profound effects on cellular responses. To better understand the role of HPKs and the signaling networks that influence influenza replication,… (more)

Subjects/Keywords: Influenza virus; host protein kinase; antiviral signaling; RNA interference; RNAi; short interfering RNA; siRNA; genome screen; microRNA; miRNA

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Andersen, L. E. (2014). MiRNA regulation and human protein kinase genes required for influenza virus replication. (Thesis). University of Georgia. Retrieved from http://hdl.handle.net/10724/27660

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):

Andersen, Lauren Elizabeth. “MiRNA regulation and human protein kinase genes required for influenza virus replication.” 2014. Thesis, University of Georgia. Accessed January 18, 2021. http://hdl.handle.net/10724/27660.

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

MLA Handbook (7^th Edition):

Andersen, Lauren Elizabeth. “MiRNA regulation and human protein kinase genes required for influenza virus replication.” 2014. Web. 18 Jan 2021.

Vancouver:

Andersen LE. MiRNA regulation and human protein kinase genes required for influenza virus replication. [Internet] [Thesis]. University of Georgia; 2014. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/10724/27660.

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

Council of Science Editors:

Andersen LE. MiRNA regulation and human protein kinase genes required for influenza virus replication. [Thesis]. University of Georgia; 2014. Available from: http://hdl.handle.net/10724/27660

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

University of Georgia

16. Adams, Allison Marie. The knockdown of Dnmt1 using small inhibitory RNA.

Degree: 2014, University of Georgia

URL: http://hdl.handle.net/10724/22045

► Increasing evidence has implicated the incomplete or aberrant reprogramming of donor nuclei as a contributing factor to the observed inefficiencies and outcomes inherent with the… (more)

Subjects/Keywords: Nuclear transfer; livestock cloning; reprogramming; genomic imprinting; DNA methylation; Dnmt1; RNA interference; RNAi; small inhibitory RNA; siRNA

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Adams, A. M. (2014). The knockdown of Dnmt1 using small inhibitory RNA. (Thesis). University of Georgia. Retrieved from http://hdl.handle.net/10724/22045

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):

Adams, Allison Marie. “The knockdown of Dnmt1 using small inhibitory RNA.” 2014. Thesis, University of Georgia. Accessed January 18, 2021. http://hdl.handle.net/10724/22045.

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

MLA Handbook (7^th Edition):

Adams, Allison Marie. “The knockdown of Dnmt1 using small inhibitory RNA.” 2014. Web. 18 Jan 2021.

Vancouver:

Adams AM. The knockdown of Dnmt1 using small inhibitory RNA. [Internet] [Thesis]. University of Georgia; 2014. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/10724/22045.

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

Council of Science Editors:

Adams AM. The knockdown of Dnmt1 using small inhibitory RNA. [Thesis]. University of Georgia; 2014. Available from: http://hdl.handle.net/10724/22045

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

17. Martins, Larissa Almeida. Efeitos da infecção por Rickettsia rickettsii sobre o perfil de expressão gênica do carrapato vetor Amblyomma cajennense.

Degree: Mestrado, Biologia da Relação Patógeno-Hospedeiro, 2014, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/42/42135/tde-13082014-201123/ ;

►

O agente etiológico da Febre Maculosa das Montanhas Rochosas (RMSF), conhecida no Brasil como Febre Maculosa Brasileira, é a bactéria Rickettsia rickettsii. Essa bactéria é… (more)

▼

O agente etiológico da Febre Maculosa das Montanhas Rochosas (RMSF), conhecida no Brasil como Febre Maculosa Brasileira, é a bactéria Rickettsia rickettsii. Essa bactéria é transmitida ao homem pela picada de diferentes espécies de carrapatos ixodídeos. No Brasil, os vetores são Amblyomma cajennense e A. aureolatum. As taxas de prevalência de R. rickettsii nas populações de carrapatos de áreas endêmicas para RMSF são baixas, em geral abaixo de 1%. Essa baixa prevalência parece estar associada a menores taxas reprodutivas e de sobrevivência de linhagens infectadas, sugerindo que R. rickettsii seja patogênica também para os seus vetores. Infecções experimentais demonstraram que 80-100% dos indivíduos de uma colônia de A. aureolatum mantida em laboratório são infectados por R. rickettsii, enquanto apenas 10-60% de A. cajennense adquirem a bactéria. Esses dados indicam que as respostas dessas duas espécies de carrapatos à infecção sejam diferentes, resultando em diferentes taxas de prevalência da bactéria. Dessa maneira, a caracterização molecular das interações entre carrapatos do gênero Amblyomma e a bactéria R. rickettsii é importante, podendo gerar informações não somente para o esclarecimento acerca dos mecanismos de patogenicidade de R. rickettsii para os carrapatos, mas também para um melhor entendimento dos mecanismos responsáveis pela aparente restringência de A. cajennense à infecção. Assim, os objetivos do presente estudo foram: (i) analisar os efeitos da infecção por R. rickettsii sobre o perfil de expressão gênica de carrapatos A. cajennense por hibridação subtrativa por supressão (SSH), (ii) validar os dados de SSH por reação em cadeia de polimerase quantitativa precedida por transcrição reversa (RT-qPCR) e (iii) caracterizar funcionalmente dois genes com expressão induzida pela infecção por RNA de interferência (RNAi). Após a análise bioinformática dos dados de SSH, 44 sequências únicas foram obtidas, das quais 36 representam genes com expressão induzida e 8 genes com expressão reprimida pela infecção. A indução dos genes codificadores da subunidade I da citocromo c oxidase (COX1), da subunidade IV da NADH desidrogenase, de uma proteína com domínio de inibidor de serina-proteases Kunitz-type (papilina-like), identificados por SSH, e de um peptídeo antimicrobiano (hebraeína), foi confirmada por RT-qPCR. O silenciamento gênico da hebraeína e da papilina-like não teve nenhum efeito na aquisição de R. rickettsii pelo vetor, indicando que, isoladamente, não são responsáveis pela proteção de A. cajennense contra a infecção. Os dados gerados pelo presente estudo abrem perspectivas para que outros genes sejam avaliados quanto ao seu papel na aquisição de R. rickettsii, os quais, no futuro, podem ser considerados como alvos para o desenvolvimento de vacinas.

The etiologic agent of the Rocky Mountain Spotted Fever (RMSF), also known as Brazilian Spotted Fever in Brazil, is the bacterium Rickettsia rickettsii. This rickettsia is transmitted to humans by the bite of various tick species. In Brazil, Amblyomma…

Advisors/Committee Members: Fogaça, Andréa Cristina.

Subjects/Keywords: Amblyomma cajennense; Amblyomma cajennense; Rickettsia rickettsii; Rickettsia rickettsii; Carrapato; Hibridação subtrativa por supressão (SSH); RNA de interferência (RNAi); RNA interference (RNAi); Suppression subtractive hybridization (SSH); Tick; Transcriptoma; Transcriptome

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Martins, L. A. (2014). Efeitos da infecção por Rickettsia rickettsii sobre o perfil de expressão gênica do carrapato vetor Amblyomma cajennense. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/42/42135/tde-13082014-201123/ ;

Chicago Manual of Style (16^th Edition):

Martins, Larissa Almeida. “Efeitos da infecção por Rickettsia rickettsii sobre o perfil de expressão gênica do carrapato vetor Amblyomma cajennense.” 2014. Masters Thesis, University of São Paulo. Accessed January 18, 2021. http://www.teses.usp.br/teses/disponiveis/42/42135/tde-13082014-201123/ ;.

MLA Handbook (7^th Edition):

Martins, Larissa Almeida. “Efeitos da infecção por Rickettsia rickettsii sobre o perfil de expressão gênica do carrapato vetor Amblyomma cajennense.” 2014. Web. 18 Jan 2021.

Vancouver:

Martins LA. Efeitos da infecção por Rickettsia rickettsii sobre o perfil de expressão gênica do carrapato vetor Amblyomma cajennense. [Internet] [Masters thesis]. University of São Paulo; 2014. [cited 2021 Jan 18]. Available from: http://www.teses.usp.br/teses/disponiveis/42/42135/tde-13082014-201123/ ;.

Council of Science Editors:

Martins LA. Efeitos da infecção por Rickettsia rickettsii sobre o perfil de expressão gênica do carrapato vetor Amblyomma cajennense. [Masters Thesis]. University of São Paulo; 2014. Available from: http://www.teses.usp.br/teses/disponiveis/42/42135/tde-13082014-201123/ ;

University of Vienna

18. Suhren, Jan Henrik. Heterochromatin components are required for limiting biogenesis of siRNAs to sequences targeted for DNA elimination in Tetrahymena.

Degree: 2015, University of Vienna

URL: http://othes.univie.ac.at/39825/

►

Heterochromatin wird allgemein als repressiver Chromatinstatus definiert. Da Heterochromatin seine eigene Ausbreitung verstärkt, müssen eukaryotische Genome diese begrenzen, um die Funktionsfähigkeit des Genoms zu gewährleisten.… (more)

▼

Heterochromatin wird allgemein als repressiver Chromatinstatus definiert. Da Heterochromatin seine eigene Ausbreitung verstärkt, müssen eukaryotische Genome diese begrenzen, um die Funktionsfähigkeit des Genoms zu gewährleisten. Dies ist besonders im Kontext der programmierten DNA Eliminierung wichtig, welche der Protist Tetrahymena thermophila nutzt, um tausende von transposonverwandten Sequenzen aus seinem somatischen Genom herauszuschneiden. Diese Sequenzen werden interne eliminierte Sequenzen (IESs) genannt. De novo Heterochromatin Etablierung ist in diesem Prozess ein essentieller Zwischenschritt, der für die Mehrzahl von IESs durch eine Population von kleinen RNAs, die sich Frühe-scnRNAs nennen, eingeleitet wird. Die übrigen IESs benötigen Späte-scnRNAs für ihre Eliminierung, welche durch eine Amplifikationsschleife produziert werden, die vermutlich auf Zyklen sich gegenseitig induzierender und verstärkender Heterochromatin Entstehung und Später-scnRNA Produktion basiert. Wie ein derartiger positiver Rückkopplungsmechanismus auf IESs begrenzt werden kann, damit ausschließlich diese eliminiert werden, ist bis heute unklar. In meiner Doktorarbeit habe ich vier Proteine identifiziert, die dafür wichtig sind, um die Späte-scnRNA Produktion auf IESs zu beschränken: das HP1-verwandte Protein Coi6p, seine zwei Interaktionspartner Coi7p und Lia5p sowie die Histone H3 Lysine 27 (H3K27) Demethylase Jmj1p. Coi6p, Coi7p und Lia5p sind Heterochromatinkomponenten. In Knockouts (KOs) von COI6, COI7 und LIA5 findet IESs Elimination größtenteils nicht statt, die Akkumulation von Kernheterochromatinkomponenten wie Pdd1p wird jedoch nicht blockiert. Dies legt nahe, dass Coi6p, Coi7p und Lia5p für die anfängliche Etablierung des Heterochromatin nicht notwendig sind, sondern in einem anderen Prozess aktiv sind. In Übereinstimmung damit konnte ich mit Hilfe der Sequenzierung von scnRNAs nachweisen, dass sich in COI6, COI7 und LIA5 KO-Zellen die Produktion von Späten-scnRNAs über die Grenzen von IESs hinaus ausbreitet. Im Gegensatz zu Coi6p, Coi7p und Lia5p findet sich in JMJ1 KO-Zellen lediglich ein schwacher Defekt bezüglich der Eliminierung von IESs. Trotzdem ist Jmj1p für die Begrenzung der Späten-scnRNA Produktion auf IESs wichtig, wie ich durch Analyse der scnRNAs in JMJ1 KO-Zellen gezeigt habe. Darüber hinaus verursacht die unpräzise Produktion von Späten-scnRNAs in COI6, COI7 und JMJ1 KO-Zellen fehlerhafte DNA Eliminierung. Zusammenfassend kann ich in der vorliegenden Arbeit zum ersten Mal zeigen, dass Heterochromatin nicht nur für die Produktion von Späten-scnRNAs wichtig ist, sondern auch für ihre IESs-Spezifität und damit für die Genauigkeit der DNA Eliminierung.

Heterochromatin is generally defined as a condensed, repressive chromatin state. Due to the self-propagating nature of heterochromatin, eukaryotic genomes must maintain heterochromatin boundaries to safeguard genome functions. This is particularly critical in the context of a process termed programmed DNA elimination, by the protist Tetrahymena thermophila…

Subjects/Keywords: 42.13 Molekularbiologie; 42.15 Zellbiologie; RNAi / RNA Interferenz / Tetrahymena / DNA Eliminierung / Heterochromatin / kleine RNAs; RNAi / RNA interference / tetrahymena / DNA elimination / heterochromatin spreading / heterochromatin boundary / small RNAs / HP1 proteins

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Suhren, J. H. (2015). Heterochromatin components are required for limiting biogenesis of siRNAs to sequences targeted for DNA elimination in Tetrahymena. (Thesis). University of Vienna. Retrieved from http://othes.univie.ac.at/39825/

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):

Suhren, Jan Henrik. “Heterochromatin components are required for limiting biogenesis of siRNAs to sequences targeted for DNA elimination in Tetrahymena.” 2015. Thesis, University of Vienna. Accessed January 18, 2021. http://othes.univie.ac.at/39825/.

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

MLA Handbook (7^th Edition):

Suhren, Jan Henrik. “Heterochromatin components are required for limiting biogenesis of siRNAs to sequences targeted for DNA elimination in Tetrahymena.” 2015. Web. 18 Jan 2021.

Vancouver:

Suhren JH. Heterochromatin components are required for limiting biogenesis of siRNAs to sequences targeted for DNA elimination in Tetrahymena. [Internet] [Thesis]. University of Vienna; 2015. [cited 2021 Jan 18]. Available from: http://othes.univie.ac.at/39825/.

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

Council of Science Editors:

Suhren JH. Heterochromatin components are required for limiting biogenesis of siRNAs to sequences targeted for DNA elimination in Tetrahymena. [Thesis]. University of Vienna; 2015. Available from: http://othes.univie.ac.at/39825/

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

University of California – Riverside

19. Kahlon, Amandeep Singh. In vitro Study of Herves Transposable Element of Anopheles gambiae and Use of RNA Interference (RNAi) in Culex quinquefasciatus.

Degree: Cell, Molecular and Developmental Biology, 2010, University of California – Riverside

URL: http://www.escholarship.org/uc/item/1d19z0z8

► Transposable elements (TEs) and RNA interference (RNAi) are excellent genetic tools that could help control the incidence and spread of mosquito-borne diseases such as malaria,… (more)

▼ Transposable elements (TEs) and RNA interference (RNAi) are excellent genetic tools that could help control the incidence and spread of mosquito-borne diseases such as malaria, dengue, yellow fever, etc. This study aimed to 1) understand the RNAi mechanism in Culex quinquefasciatus, which will be used to study genes involved in pathogen transmission and will help reveal role of RNAi in antiviral immunity in this species; and 2) characterize the DNA sequences that regulate Herves transposase binding, which will help us understand its pre- and post-integration behavior within the host. By using the white eye-pigmentation gene as a marker for RNAi function we demonstrated that introducing dsRNA into embryos of Cx. quinquefasciatus induces a specific functional RNAi response, which silenced the white gene, consequently allowing a white-eye phenotype to be produced in the hatched larvae and adults. Sequence-specific knockdown of key RNAi components was achieved by introducing the homologous dsRNA into embryos of Cx. quinquefasciatus. We found ago2-mediated slicing to be more critical than dcr2-mediated dicing for the functional RNAi response in Cx. quinquefasciatus. Our phylogenetic analysis confirmed the previously reported results that RNAi components for small RNA biogenesis are present in Cx. quinquefasciatus. In addition, we found important differences in the number and the expression of ago genes, the predicted domain architecture of Dcr, and the RNAi response between Cx. quinquefasciatus and the model organism D. melanogaster. Transposition of Class II TEs is regulated by both cis-acting sequences and trans-acting host factors. In this study, we used purified Herves transposase to characterize the specific DNA-binding sites of the Herves transposase. The purified active Herves transposase showed site-specific binding to the subterminal and terminal sequences of the L- and R- ends of the element, respectively. Furthermore, the transposase bound strongly with the R-TIR but failed to bind to the L-TIR. We identified an 8bp sequence repeat as the transposase binding motif that is conserved on both the L- and R-end sequences and is critical and sufficient for Herves transposase binding.

Subjects/Keywords: Biology, Molecular; Biology, Genetics; Biology, Entomology; Anopheles gambiae; Culex quinquefascaitus; Herves; RNAi; RNA Interference; Transposable Element

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Kahlon, A. S. (2010). In vitro Study of Herves Transposable Element of Anopheles gambiae and Use of RNA Interference (RNAi) in Culex quinquefasciatus. (Thesis). University of California – Riverside. Retrieved from http://www.escholarship.org/uc/item/1d19z0z8

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):

Kahlon, Amandeep Singh. “In vitro Study of Herves Transposable Element of Anopheles gambiae and Use of RNA Interference (RNAi) in Culex quinquefasciatus.” 2010. Thesis, University of California – Riverside. Accessed January 18, 2021. http://www.escholarship.org/uc/item/1d19z0z8.

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

MLA Handbook (7^th Edition):

Kahlon, Amandeep Singh. “In vitro Study of Herves Transposable Element of Anopheles gambiae and Use of RNA Interference (RNAi) in Culex quinquefasciatus.” 2010. Web. 18 Jan 2021.

Vancouver:

Kahlon AS. In vitro Study of Herves Transposable Element of Anopheles gambiae and Use of RNA Interference (RNAi) in Culex quinquefasciatus. [Internet] [Thesis]. University of California – Riverside; 2010. [cited 2021 Jan 18]. Available from: http://www.escholarship.org/uc/item/1d19z0z8.

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

Council of Science Editors:

Kahlon AS. In vitro Study of Herves Transposable Element of Anopheles gambiae and Use of RNA Interference (RNAi) in Culex quinquefasciatus. [Thesis]. University of California – Riverside; 2010. Available from: http://www.escholarship.org/uc/item/1d19z0z8

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

Universidade Estadual de Campinas

20. Araujo, Patricia Aline Oliveira Ribeiro de Aguiar. Analise de expressão do gene Lgi1 durante o desenvolvimento do sistema nervoso central e seu silenciamento utilizando a tecnica de interferencia por RNA: Lgi1 gene expression analysis during the central nervous system development and its silencing using the interference RNA.

Degree: 2008, Universidade Estadual de Campinas

URL: http://repositorio.unicamp.br/jspui/handle/REPOSIP/309740

► Abstract: Introduction/Objectives: Mutations in the LGI1 gene were described in some patients with autosomal dominant partial epilepsy with auditory features and preliminary functional studies point… (more)

▼ Abstract: Introduction/Objectives: Mutations in the LGI1 gene were described in some patients with autosomal dominant partial epilepsy with auditory features and preliminary functional studies point to a possible involvement of LGI1 with migration and/or neuronal proliferation. However, the precise function of LGI1 remains unknown. The objective of the present study was to determine the expression pattern of the Lgi1 gene in mice brain during central nervous system (CNS) development and in adult animals. In addition, we aimed to silence Lgi1 in mouse brain using RNA interference (RNAi). Methods: Programmed mating was carried with Balb/c mice in order to obtain embryos of different ages. The brains of three animals at the following ages were removed and the right and left hemispheres were separated: E15, E17, E18 days (E: embryo days), P1, P7, P14 (P: post-natal days), 4, 6, 8 and 24 weeks old. We also studied E13 whole head animals. In addition we studied three different regions from 5 weeks-old animal brains: neocortex, hippocampus and cerebellum. Gene expression assays were carried out using real time PCR with the TaqMan® system and western blot experiments. RNAi was performed using different methods for injection of interfering molecules into the neonate and adult brains. We also used different molecule designs and concentrations, as well as the number and the local of injections was varied. Furthermore, we performed in vitro RNAi experiments using a glioblastoma cell line, U138MG, and a murine cell line, Neuro2a. Gene silencing confirmation was carried out by real time PCR and western blot assays. Results: Lgi1 gene expression was significantly low during the intrauterine ages increasing progressively until the adult stages. Samples from adult animals presented a 35 fold increase in expression as compared to E13 samples, using Gapdh as endogenous control, and when we use ?-actin, adult samples presented approximately 28 fold increase in Lgi1 expression. There were no statistical differences between Lgi1 gene expression test between right and left hemispheres. However, a significant difference in expression was found among the different ages studied. The western blot showed higher expression of the Lgi1 protein in the most advanced ages analyzed, confirming the expression profile observed in the real time PCR studies. However, we did not find any statistic difference between the three regions of the brain studied. In addition, we achieved significant gene silencing of Lgi1, reduction of expression of approximately 50%, in brain of adult animals using RNAi and the local electroporation method. In addition, we demonstrated up to 99% silencing of LGI1 in cell culture. Conclusions/Discussion: The Lgi1 expression profile, which is characterized by low expression in the initial stages of development with progressive increase as the animal developed, could be explained by a possible inhibitory functional role in neuronal proliferation during CNS development. Lgi1 gene silencing in adult brain using RNAi technique was… Advisors/Committee Members: UNIVERSIDADE ESTADUAL DE CAMPINAS (CRUESP), Lopes-Cendes, Íscia Teresinha, 1964- (advisor), Universidade Estadual de Campinas. Faculdade de Ciências Médicas (institution), Programa de Pós-Graduação em Fisiopatologia Médica (nameofprogram), Bittencourt, Jackson Cioni (committee member), Neto, Mario Pedrazzoli (committee member), Melo, Mônica Barbosa de (committee member), Sonati, Maria de Fátima (committee member).

Subjects/Keywords: Epilepsia; RNAi; Neurogenética; Epilepsy; RNA interference; Neurogenetics

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Araujo, P. A. O. R. d. A. (2008). Analise de expressão do gene Lgi1 durante o desenvolvimento do sistema nervoso central e seu silenciamento utilizando a tecnica de interferencia por RNA: Lgi1 gene expression analysis during the central nervous system development and its silencing using the interference RNA. (Thesis). Universidade Estadual de Campinas. Retrieved from http://repositorio.unicamp.br/jspui/handle/REPOSIP/309740

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):

Araujo, Patricia Aline Oliveira Ribeiro de Aguiar. “Analise de expressão do gene Lgi1 durante o desenvolvimento do sistema nervoso central e seu silenciamento utilizando a tecnica de interferencia por RNA: Lgi1 gene expression analysis during the central nervous system development and its silencing using the interference RNA.” 2008. Thesis, Universidade Estadual de Campinas. Accessed January 18, 2021. http://repositorio.unicamp.br/jspui/handle/REPOSIP/309740.

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

MLA Handbook (7^th Edition):

Araujo, Patricia Aline Oliveira Ribeiro de Aguiar. “Analise de expressão do gene Lgi1 durante o desenvolvimento do sistema nervoso central e seu silenciamento utilizando a tecnica de interferencia por RNA: Lgi1 gene expression analysis during the central nervous system development and its silencing using the interference RNA.” 2008. Web. 18 Jan 2021.

Vancouver:

Araujo PAORdA. Analise de expressão do gene Lgi1 durante o desenvolvimento do sistema nervoso central e seu silenciamento utilizando a tecnica de interferencia por RNA: Lgi1 gene expression analysis during the central nervous system development and its silencing using the interference RNA. [Internet] [Thesis]. Universidade Estadual de Campinas; 2008. [cited 2021 Jan 18]. Available from: http://repositorio.unicamp.br/jspui/handle/REPOSIP/309740.

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

Council of Science Editors:

Araujo PAORdA. Analise de expressão do gene Lgi1 durante o desenvolvimento do sistema nervoso central e seu silenciamento utilizando a tecnica de interferencia por RNA: Lgi1 gene expression analysis during the central nervous system development and its silencing using the interference RNA. [Thesis]. Universidade Estadual de Campinas; 2008. Available from: http://repositorio.unicamp.br/jspui/handle/REPOSIP/309740

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

University of Edinburgh

21. Marr, Edward John. RNA interference (RNAi) for selective gene silencing in Astigmatid mites.

Degree: PhD, 2016, University of Edinburgh

URL: http://hdl.handle.net/1842/25722

► Psoroptic mange, caused by the Astigmatid mite Psoroptes ovis, is an ectoparasitic disease of significant economic importance to agriculture on a global scale and poses… (more)

Subjects/Keywords: 572.8; RNA interference; RNAi; ectoparasit; mite; gene silencing; microRNA; Astigmatid; Psoroptes ovis; Sarcoptes scabiei; Dermatophagoides pteronyssinus

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Marr, E. J. (2016). RNA interference (RNAi) for selective gene silencing in Astigmatid mites. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/25722

Chicago Manual of Style (16^th Edition):

Marr, Edward John. “RNA interference (RNAi) for selective gene silencing in Astigmatid mites.” 2016. Doctoral Dissertation, University of Edinburgh. Accessed January 18, 2021. http://hdl.handle.net/1842/25722.

MLA Handbook (7^th Edition):

Marr, Edward John. “RNA interference (RNAi) for selective gene silencing in Astigmatid mites.” 2016. Web. 18 Jan 2021.

Vancouver:

Marr EJ. RNA interference (RNAi) for selective gene silencing in Astigmatid mites. [Internet] [Doctoral dissertation]. University of Edinburgh; 2016. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/1842/25722.

Council of Science Editors:

Marr EJ. RNA interference (RNAi) for selective gene silencing in Astigmatid mites. [Doctoral Dissertation]. University of Edinburgh; 2016. Available from: http://hdl.handle.net/1842/25722

22. TANG HOCK CHUN. Characterisation of agrobacterium VirD2 interacting protein DIP and its homologues.

Degree: 2006, National University of Singapore

URL: http://scholarbank.nus.edu.sg/handle/10635/27850

Subjects/Keywords: Agrobacterium tumefaciens; VirD2-interacting protein(DIP); RNA interference(RNAi)

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

CHUN, T. H. (2006). Characterisation of agrobacterium VirD2 interacting protein DIP and its homologues. (Thesis). National University of Singapore. Retrieved from http://scholarbank.nus.edu.sg/handle/10635/27850

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):

CHUN, TANG HOCK. “Characterisation of agrobacterium VirD2 interacting protein DIP and its homologues.” 2006. Thesis, National University of Singapore. Accessed January 18, 2021. http://scholarbank.nus.edu.sg/handle/10635/27850.

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

MLA Handbook (7^th Edition):

CHUN, TANG HOCK. “Characterisation of agrobacterium VirD2 interacting protein DIP and its homologues.” 2006. Web. 18 Jan 2021.

Vancouver:

CHUN TH. Characterisation of agrobacterium VirD2 interacting protein DIP and its homologues. [Internet] [Thesis]. National University of Singapore; 2006. [cited 2021 Jan 18]. Available from: http://scholarbank.nus.edu.sg/handle/10635/27850.

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

Council of Science Editors:

CHUN TH. Characterisation of agrobacterium VirD2 interacting protein DIP and its homologues. [Thesis]. National University of Singapore; 2006. Available from: http://scholarbank.nus.edu.sg/handle/10635/27850

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

Kansas State University

23. Aguirre Rojas, Lina Maria. Towards the development of soybean resistance to Dectes texanus LeConte (Coleoptera: Cerambycidae): evaluation of conventional soybean resistance in the soybean plant introduction 165673, transcriptomic analyses, and gene silencing by RNA interference.

Degree: PhD, Department of Entomology, 2019, Kansas State University

URL: http://hdl.handle.net/2097/39649

► Dectes texanus (Coleoptera:Cerambyciade) larvae devastate soybeans in the U. S. midwestern states by girdling and tunneling inside the stems. Reduction of natural sunflower and ragweed… (more)

▼ Dectes texanus (Coleoptera:Cerambyciade) larvae devastate soybeans in the U. S. midwestern states by girdling and tunneling inside the stems. Reduction of natural sunflower and ragweed hosts may have promoted the use soybean as a host since the late 1950s. Development of soybean varieties resistant to D. texanus is of importance to manage this pest since harvesting as soon as possible is the only option available to farmers to reduce yield losses. The soybean plant introduction (PI) 165673 reduces the number of D. texanus larvae, but survivors are still found at 21 d post infestation that may damage and girdle the PI165673 stems at the end of the growing season. Soybean resistance to D. texanus can be enhanced with the delivery of double stranded RNA (dsRNA) in planta to suppress gene expression by RNA interference (RNAi). DsRNA expressed in planta can be designed specifically to target and silence D. texanus genes important for development. D. texanus genes uniquely or highly expressed when fed soybean compared to those fed their natural hosts or artificial diet can be exploited to impair the development of D. texanus with RNAi. Although, dsRNA-based silencing is successful in other cerambycids, its use with D. texanus is untested. The objectives of this dissertation are: 1) to evaluate stem girdling and tunneling by D. texanus in soybean PI165673 at the end of the 2014 growing season in Kansas; 2) to compare the transcriptomes of D. texanus larvae fed soybean, wild sunflower, giant ragweed or artificial diet; and 3) to silence the Laccase2 (Lac2) and Chitin synthase2 (CHS2) genes in D. texanus by feeding larvae artificial diet coated with Lac2 or CHS2-dsRNA. The stems of infested PI165673 had 20% less tunneling by D. texanus larvae compared to the susceptible control K07_1544 at the end of the growing season in 2014 in Kansas. However, this result may be overestimated since the PI16563 plants had not reached full maturity and development compared to K07_1544. Therefore, assessment of stem tunneling (%) and girdling damage should be conducted in Southern States where the PI165637 can finish their development. The PI165673 resistance negatively affected the development and survival of D. texanus first instar larvae, but surviving larvae developed until the last instar stage before the end of the growing season in Kansas. These larvae can potentially gridle the PI165673 plants. A D. texanus de novo transcriptome was assembled for differential gene expression analyses. Five and six unigenes were commonly up-regulated and down-regulated in K07_1544-fed larvae compared to those fed wild sunflower, giant ragweed, or artificial diet (FDR < 0.05, fold change cut off ≥ ± 1.5). Unigenes coding for a lipocalin, an ecdysteroid kinase, and a major facilitator transporter were among the commonly up-regulated unigenes in soybean-fed larvae. Unigenes coding for two insect cuticle proteins, a glycosyl hydrolase 45, a transglutaminase, and a chitin binding peritrophin-A domain were among the commonly down-regulated unigenes in… Advisors/Committee Members: C. Michael Smith.

Subjects/Keywords: Dectes texanus; Host plant resistance; De novo transcriptome; Differential expression analysis; Gene silencing; RNA interference RNAi

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Aguirre Rojas, L. M. (2019). Towards the development of soybean resistance to Dectes texanus LeConte (Coleoptera: Cerambycidae): evaluation of conventional soybean resistance in the soybean plant introduction 165673, transcriptomic analyses, and gene silencing by RNA interference. (Doctoral Dissertation). Kansas State University. Retrieved from http://hdl.handle.net/2097/39649

Chicago Manual of Style (16^th Edition):

Aguirre Rojas, Lina Maria. “Towards the development of soybean resistance to Dectes texanus LeConte (Coleoptera: Cerambycidae): evaluation of conventional soybean resistance in the soybean plant introduction 165673, transcriptomic analyses, and gene silencing by RNA interference.” 2019. Doctoral Dissertation, Kansas State University. Accessed January 18, 2021. http://hdl.handle.net/2097/39649.

MLA Handbook (7^th Edition):

Aguirre Rojas, Lina Maria. “Towards the development of soybean resistance to Dectes texanus LeConte (Coleoptera: Cerambycidae): evaluation of conventional soybean resistance in the soybean plant introduction 165673, transcriptomic analyses, and gene silencing by RNA interference.” 2019. Web. 18 Jan 2021.

Vancouver:

Aguirre Rojas LM. Towards the development of soybean resistance to Dectes texanus LeConte (Coleoptera: Cerambycidae): evaluation of conventional soybean resistance in the soybean plant introduction 165673, transcriptomic analyses, and gene silencing by RNA interference. [Internet] [Doctoral dissertation]. Kansas State University; 2019. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/2097/39649.

Council of Science Editors:

Aguirre Rojas LM. Towards the development of soybean resistance to Dectes texanus LeConte (Coleoptera: Cerambycidae): evaluation of conventional soybean resistance in the soybean plant introduction 165673, transcriptomic analyses, and gene silencing by RNA interference. [Doctoral Dissertation]. Kansas State University; 2019. Available from: http://hdl.handle.net/2097/39649

Texas A&M University

24. Wang, Tao. Characterization of AtSUVR3 functions in Arabidopsis thaliana using RNA interference.

Degree: PhD, Biology, 2009, Texas A&M University

URL: http://hdl.handle.net/1969.1/ETD-TAMU-1427

► Variability of transgene expression levels resulting from gene silencing is considered as ahindrance to the successful application of plant genetic engineering. Towards alleviatinggene silencing, I… (more)

▼ Variability of transgene expression levels resulting from gene silencing is considered as ahindrance to the successful application of plant genetic engineering. Towards alleviatinggene silencing, I decided to screen for novel genes involved in transgene silencing and toinvestigate how these genes regulate plant development. Genes encoding putative chromatinremodeling factors, especially those including a SET domain, were selected as candidatetargets. A bioinformatic analysis of the Arabidopsis SET genes (AtSET) was performed andthese genes were classified into 6 groups based on the domain architecture. RNA interference (RNAi) vectors were constructed for ~ 20 AtSET genes and wereintroduced into both wild type lines and transgenic lines silenced for a GFP reporter gene.Surprisingly, altered developmental phenotypes were only observed for three constructs,raising questions as to the effectiveness of the RNAi approach for the chosen Arabidopsissystem. To assess this situation, I targeted a phytoene desaturase (PDS) gene using the sameRNAi approach. Inactivation of PDS renders plant a readily identifiable phenotype. Whereasthe RNAi penetrance in Arabidopsis can be very high, the expressivity of RNAi in varioustissues and among different plants can vary dramatically. Contradictory to previous reports,I found that there is correlation between transcript level and silencing phenotype. Possiblereasons for this discrepancy are discussed. No apparent correlation between transgene copynumber and RNAi phenotypes was observed. Among the three RNAi constructs that caused an abnormal development inArabidopsis, K-23 which targets SuvR3 has the highest expressivity and could reactivate asilenced GFP locus. SuvR3 RNAi lines were selfed for six generations and were screenedfor morphological phenotypes. Abnormal number of flower organs, loss of viability of malegametophytes, and decreased seedling germination percentage were found in SuvR3 RNAilines. A progressive increase in both severity and frequency of abnormal phenotypes wereseen in subsequent generations, suggesting an epigenetic regulatory mechanism involvedwith SuvR3. Alternative splicing of SuvR3 was also observed in most of Arabidopsis tissues.One of the protein isoforms, SuvR3, lacks 16 amino acids within the highly conserved SETdomain. Possible effects of isoform interaction are proposed. Advisors/Committee Members: Hall, Timothy C (advisor), Chen, Jeffrey Z (committee member), McKnight, Thomas D (committee member), Pepper, Alan E (committee member).

Subjects/Keywords: RNA interference; RNAi penetrance; RNAi expressivity; gene silencing; SET domain; alternative splicing

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Wang, T. (2009). Characterization of AtSUVR3 functions in Arabidopsis thaliana using RNA interference. (Doctoral Dissertation). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-1427

Chicago Manual of Style (16^th Edition):

Wang, Tao. “Characterization of AtSUVR3 functions in Arabidopsis thaliana using RNA interference.” 2009. Doctoral Dissertation, Texas A&M University. Accessed January 18, 2021. http://hdl.handle.net/1969.1/ETD-TAMU-1427.

MLA Handbook (7^th Edition):

Wang, Tao. “Characterization of AtSUVR3 functions in Arabidopsis thaliana using RNA interference.” 2009. Web. 18 Jan 2021.

Vancouver:

Wang T. Characterization of AtSUVR3 functions in Arabidopsis thaliana using RNA interference. [Internet] [Doctoral dissertation]. Texas A&M University; 2009. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-1427.

Council of Science Editors:

Wang T. Characterization of AtSUVR3 functions in Arabidopsis thaliana using RNA interference. [Doctoral Dissertation]. Texas A&M University; 2009. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-1427

Harvard University

25. Bhatia, Sonya. Characterization of Systemic RNAi Mutants in Caenorhabditis Elegans.

Degree: PhD, 2019, Harvard University

URL: http://nrs.harvard.edu/urn-3:HUL.InstRepos:42106929

►

RNA interference (RNAi) is sequence-specific gene silencing triggered by double-stranded (ds)RNA. When dsRNA is expressed or introduced into one cell and is transported to and… (more)

▼

RNA interference (RNAi) is sequence-specific gene silencing triggered by double-stranded (ds)RNA. When dsRNA is expressed or introduced into one cell and is transported to and initiates RNAi in other cells, it is called systemic RNAi. Systemic RNAi enables easy and efficient whole-animal gene silencing in C. elegans. While the specificity and potency of RNAi offers tremendous therapeutic potential to treat genetic and infectious disease, effective delivery to human cells remains a major challenge. Understanding systemic RNAi transport pathways in C. elegans may help identify strategies for effective delivery in mammals. The Hunter lab conducted a genetic screen in C. elegans that identified five genes, sid-1 through -5, that are specifically required for systemic RNAi. Using whole genome sequencing, I identified sid-4 as nck-1, a non-receptor tyrosine kinase adaptor protein. Interestingly, sid-3 is homologous to ACK tyrosine kinases that physically interact with NCK homologs. SID-3 and SID-4 are both broadly expressed and show similar subcellular localization patterns. My analysis showed that sid-4, like sid-3, is a dose-dependent import mutant. Although sid-3 and sid-4 mutants showed similar patterns of RNAi resistance, the sid-3; sid-4 double mutant was significantly more RNAi defective than either single mutant, indicating that these proteins do not function only in a simple linear pathway. Consistent with their identity as conserved signaling proteins, their activity is required to mediate temperature-dependent systemic RNAi efficiency effects. Interestingly, both mutants form dauers at elevated temperatures (27°C), suggesting that RNA mobility may regulate growth and development. A likely model is that SID-3/SID-4 act to modulate the activity of the conserved dsRNA conducting channel, SID-1.

Medical Sciences

Advisors/Committee Members: Heiman, Maxwell (advisor), Blackwell, T. Keith (committee member), Apfeld, Javier (committee member), Cram, Erin (committee member).

Subjects/Keywords: RNAi; systemic RNAi; sid-4; sid-3; dsRNA; RNA interference; NCK; Caenorhabditis elegans; C. elegans; dauer; ACK; adaptor protein; tyrosine kinase; ver-1; receptor tyrosine kinase

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Bhatia, S. (2019). Characterization of Systemic RNAi Mutants in Caenorhabditis Elegans. (Doctoral Dissertation). Harvard University. Retrieved from http://nrs.harvard.edu/urn-3:HUL.InstRepos:42106929

Chicago Manual of Style (16^th Edition):

Bhatia, Sonya. “Characterization of Systemic RNAi Mutants in Caenorhabditis Elegans.” 2019. Doctoral Dissertation, Harvard University. Accessed January 18, 2021. http://nrs.harvard.edu/urn-3:HUL.InstRepos:42106929.

MLA Handbook (7^th Edition):

Bhatia, Sonya. “Characterization of Systemic RNAi Mutants in Caenorhabditis Elegans.” 2019. Web. 18 Jan 2021.

Vancouver:

Bhatia S. Characterization of Systemic RNAi Mutants in Caenorhabditis Elegans. [Internet] [Doctoral dissertation]. Harvard University; 2019. [cited 2021 Jan 18]. Available from: http://nrs.harvard.edu/urn-3:HUL.InstRepos:42106929.

Council of Science Editors:

Bhatia S. Characterization of Systemic RNAi Mutants in Caenorhabditis Elegans. [Doctoral Dissertation]. Harvard University; 2019. Available from: http://nrs.harvard.edu/urn-3:HUL.InstRepos:42106929

Virginia Tech

26. Bhattacharjee, Puranjoy. Correlation Between Computed Equilibrium Secondary Structure Free Energy and siRNA Efficiency.

Degree: MS, Computer Science, 2009, Virginia Tech

URL: http://hdl.handle.net/10919/34643

► We have explored correlations between the measured efficiency of the RNAi process and several computed signatures that characterize equilibrium secondary structure of the partic- ipating… (more)

Subjects/Keywords: RNAi efficiency; RNA interference(RNAi); RNAi equilibrium thermodynamics; Support Vector Machine; RNA secondary structure

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Bhattacharjee, P. (2009). Correlation Between Computed Equilibrium Secondary Structure Free Energy and siRNA Efficiency. (Masters Thesis). Virginia Tech. Retrieved from http://hdl.handle.net/10919/34643

Chicago Manual of Style (16^th Edition):

Bhattacharjee, Puranjoy. “Correlation Between Computed Equilibrium Secondary Structure Free Energy and siRNA Efficiency.” 2009. Masters Thesis, Virginia Tech. Accessed January 18, 2021. http://hdl.handle.net/10919/34643.

MLA Handbook (7^th Edition):

Bhattacharjee, Puranjoy. “Correlation Between Computed Equilibrium Secondary Structure Free Energy and siRNA Efficiency.” 2009. Web. 18 Jan 2021.

Vancouver:

Bhattacharjee P. Correlation Between Computed Equilibrium Secondary Structure Free Energy and siRNA Efficiency. [Internet] [Masters thesis]. Virginia Tech; 2009. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/10919/34643.

Council of Science Editors:

Bhattacharjee P. Correlation Between Computed Equilibrium Secondary Structure Free Energy and siRNA Efficiency. [Masters Thesis]. Virginia Tech; 2009. Available from: http://hdl.handle.net/10919/34643

Universidade Estadual de Campinas

27. Pereira, Tiago Campos. Estudo de possiveis aplicações médicas da interferencia por RNA: RNA interference: studies on possible medical applications.

Degree: 2005, Universidade Estadual de Campinas

URL: http://repositorio.unicamp.br/jspui/handle/REPOSIP/316861

Subjects/Keywords: RNAi; RNA interferente pequeno; Inativação gênica; Interferência de RNA; RNAi; Small interfering RNA; Gene silencing; RNA interference

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Pereira, T. C. (2005). Estudo de possiveis aplicações médicas da interferencia por RNA: RNA interference: studies on possible medical applications. (Thesis). Universidade Estadual de Campinas. Retrieved from http://repositorio.unicamp.br/jspui/handle/REPOSIP/316861

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):

Pereira, Tiago Campos. “Estudo de possiveis aplicações médicas da interferencia por RNA: RNA interference: studies on possible medical applications.” 2005. Thesis, Universidade Estadual de Campinas. Accessed January 18, 2021. http://repositorio.unicamp.br/jspui/handle/REPOSIP/316861.

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

MLA Handbook (7^th Edition):

Pereira, Tiago Campos. “Estudo de possiveis aplicações médicas da interferencia por RNA: RNA interference: studies on possible medical applications.” 2005. Web. 18 Jan 2021.

Vancouver:

Pereira TC. Estudo de possiveis aplicações médicas da interferencia por RNA: RNA interference: studies on possible medical applications. [Internet] [Thesis]. Universidade Estadual de Campinas; 2005. [cited 2021 Jan 18]. Available from: http://repositorio.unicamp.br/jspui/handle/REPOSIP/316861.

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

Council of Science Editors:

Pereira TC. Estudo de possiveis aplicações médicas da interferencia por RNA: RNA interference: studies on possible medical applications. [Thesis]. Universidade Estadual de Campinas; 2005. Available from: http://repositorio.unicamp.br/jspui/handle/REPOSIP/316861

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

University of Iowa

28. Kim, NaJung. Rationale design of polymeric siRNA delivery systems.

Degree: PhD, Biomedical Engineering, 2011, University of Iowa

URL: https://ir.uiowa.edu/etd/1237

► Regulation of gene expression using small interfering RNA (siRNA) is a promising strategy for research and treatment of numerous diseases. However, siRNA cannot easily… (more)

▼ Regulation of gene expression using small interfering RNA (siRNA) is a promising strategy for research and treatment of numerous diseases. However, siRNA cannot easily cross the cell membrane due to its inherent instability, large molecular weight and anionic nature. For this reason, a carrier that protects, delivers and unloads siRNA is required for successful gene silencing. The goal of this research was to develop a potential siRNA delivery system for in vitro and in vivo applications using cationic polymers, chitosan and polyethylenimine (PEI), poly(ethylene glycol) (PEG), mannose, and poly(D,L-lactic-co-glycolic acid) (PLGA). Furthermore, the delivery system was constructed in two different ways to explore the effect of mannose location in the structure. In the first approach, mannose and PEG were directly conjugated to the chitosan/PEI backbone, while mannose was connected to the chitosan/PEI backbone through PEG spacer in the second approach. First, the ability of modified chitosan polymers to complex and deliver siRNA for gene silencing was investigated. Despite the modified chitosan polymers successfully formed nanoplexes with siRNA, entered target cells and reduced cytotoxicity of unmodified chitosan, they showed limited gene silencing efficiency. For this reason, modified PEIs were examined to improve in vitro gene knockdown. The modified PEI polymers also complexed with siRNA and facilitated endocytosis of the nanoplexes. In addition, the modifications reduced inherent cytotoxicity of unmodified PEI without compromising the gene silencing efficiency on both mRNA and protein levels. Interestingly, we found that complexation of siRNA with PEI-PEG-mannose resulted in higher cell uptake and gene silencing than complexes made with mannose-PEI-PEG. Finally, the effect of sustained release of the mannosylated pegylated PEI/siRNA nanoplexes on gene silencing was tested by encapsulating the nanoplexes within PLGA microparticles. The modified PEIs enhanced the entrapment efficiency of siRNA into the particles and resulted in reduced initial burst followed by sustained release. Incorporating the modified PEIs increased cellular uptake of siRNA, whereas it did not enhance in vitro gene knockdown efficiency due to the sustained release properties. The modified PEIs reduced the in vitro cytotoxicity and in vivo hepatotoxicity of the PLGA microparticles. In addition, encapsulating the nanoplexes into PLGA microparticles further reduced the cytotoxicity of PEI. Throughout the study, the second structure was proven more efficacious than the first structure in cellular uptake, gene silencing, siRNA encapsulation, and sustained release. We have developed novel polymeric siRNA delivery systems that enhance delivery efficiency and cellular uptake of siRNA. They have great potential for utility as a long-acting siRNA delivery system in biomedical research. Advisors/Committee Members: Salem, Aliasger K. (supervisor).

Subjects/Keywords: chitosan; gene delivery; poly(D,L-lactic-co-glycolic acid); polyethylenimine; RNA interference (RNAi); small interfering RNA (siRNA); Biomedical Engineering and Bioengineering

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Kim, N. (2011). Rationale design of polymeric siRNA delivery systems. (Doctoral Dissertation). University of Iowa. Retrieved from https://ir.uiowa.edu/etd/1237

Chicago Manual of Style (16^th Edition):

Kim, NaJung. “Rationale design of polymeric siRNA delivery systems.” 2011. Doctoral Dissertation, University of Iowa. Accessed January 18, 2021. https://ir.uiowa.edu/etd/1237.

MLA Handbook (7^th Edition):

Kim, NaJung. “Rationale design of polymeric siRNA delivery systems.” 2011. Web. 18 Jan 2021.

Vancouver:

Kim N. Rationale design of polymeric siRNA delivery systems. [Internet] [Doctoral dissertation]. University of Iowa; 2011. [cited 2021 Jan 18]. Available from: https://ir.uiowa.edu/etd/1237.

Council of Science Editors:

Kim N. Rationale design of polymeric siRNA delivery systems. [Doctoral Dissertation]. University of Iowa; 2011. Available from: https://ir.uiowa.edu/etd/1237

University of Western Ontario

29. Di Cresce, Christine. siRNA Targeting of Thymidylate Synthase, Thymidine Kinase 1 and Thymidine Kinase 2 as an Anticancer Therapy: A Combinatorial RNAi Approach.

Degree: 2014, University of Western Ontario

URL: https://ir.lib.uwo.ca/etd/2032

► Thymidylate synthase (TS) is the only de novo source of thymidylate (dTMP) for DNA synthesis and repair. Drugs targeting TS protein are a mainstay in… (more)

▼ Thymidylate synthase (TS) is the only de novo source of thymidylate (dTMP) for DNA synthesis and repair. Drugs targeting TS protein are a mainstay in cancer treatment but off-target effects and toxicity limit their use. Cytosolic thymidine kinase (TK1) and mitochondrial thymidine kinase (TK2) contribute to an alternative dTMP-producing pathway, by salvaging thymidine from the tumour milieu, and may modulate resistance to TS-targeting drugs. We have previously shown that TS antisense molecules (oligodeoxynucleotides, ODNs, and small interfering siRNA, siRNA) sensitize tumour cells, both in vitro and in vivo, to TS targeting drugs. As both TS and TKs contribute to cellular dTMP, we hypothesized that TKs mediate resistance to the capacity of TS siRNA to sensitize tumour cells to TS-targeting drugs. Downregulation of TKs with siRNA enhanced the capacity of TS siRNA to sensitize tumour cells to traditional TS protein-targeting drugs (5FUdR and pemetrexed). Combined downregulation of these enzymes is an attractive strategy to enhance TS-targeted anticancer therapy. TK2 can phosphorylate both thymidine and deoxycytidine to generate dTMP and dCMP, precursors for dTTP and dCTP, respectively. dCTP negatively regulates deoxycytidine kinase (dCK), another enzyme that phosphorylates deoxycytidine as well as the anticancer drug gemcitabine. Antisense knockdown of TK2 could reduce TK2-produced dCMP, thus decreasing dCTP levels and inhibition of dCK, and lead to increased dCK activity, gemcitabine activation, and anticancer effectiveness. Given the substrate promiscuity of TK2, we hypothesized that: (1) TK2 can mediate human tumour cell resistance to gemcitabine, (2) antisense downregulation of TK2 can overcome that resistance, and (3) TK2 siRNA-induced drug sensitization results in mitochondrial damage. siRNA downregulation of TK2 expression sensitized MCF7 and HeLa cells to gemcitabine, but did not sensitize A549 cells (low TK2 expresser). Treatment with TK2 siRNA and gemcitabine: 1) decreased mitochondrial redox status, 2) decreased mitochondrial DNA (mtDNA:nDNA ratio), and 3) decreased mitochondrial activity. This is the first demonstration of a direct role for TK2 in gemcitabine resistance, or any independent role in cancer drug resistance, and further distinguishes TK2 from other dTMP-producing enzymes.

Subjects/Keywords: small interfering RNA (siRNA); RNA interference (RNAi); combinatorial RNAi; thymidylate synthase (TS); thymidine kinase 1 (TK1); thymidine kinase 2 (TK2); cancer; anticancer drugs; gemcitabine; deoxycytidine kinase (dCK); anticancer therapy; mitochondrial DNA; mitochondrial activity; Biology; Cancer Biology; Molecular Biology; Pharmacology

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Di Cresce, C. (2014). siRNA Targeting of Thymidylate Synthase, Thymidine Kinase 1 and Thymidine Kinase 2 as an Anticancer Therapy: A Combinatorial RNAi Approach. (Thesis). University of Western Ontario. Retrieved from https://ir.lib.uwo.ca/etd/2032

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):

Di Cresce, Christine. “siRNA Targeting of Thymidylate Synthase, Thymidine Kinase 1 and Thymidine Kinase 2 as an Anticancer Therapy: A Combinatorial RNAi Approach.” 2014. Thesis, University of Western Ontario. Accessed January 18, 2021. https://ir.lib.uwo.ca/etd/2032.

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

MLA Handbook (7^th Edition):

Di Cresce, Christine. “siRNA Targeting of Thymidylate Synthase, Thymidine Kinase 1 and Thymidine Kinase 2 as an Anticancer Therapy: A Combinatorial RNAi Approach.” 2014. Web. 18 Jan 2021.

Vancouver:

Di Cresce C. siRNA Targeting of Thymidylate Synthase, Thymidine Kinase 1 and Thymidine Kinase 2 as an Anticancer Therapy: A Combinatorial RNAi Approach. [Internet] [Thesis]. University of Western Ontario; 2014. [cited 2021 Jan 18]. Available from: https://ir.lib.uwo.ca/etd/2032.

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

Council of Science Editors:

Di Cresce C. siRNA Targeting of Thymidylate Synthase, Thymidine Kinase 1 and Thymidine Kinase 2 as an Anticancer Therapy: A Combinatorial RNAi Approach. [Thesis]. University of Western Ontario; 2014. Available from: https://ir.lib.uwo.ca/etd/2032

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

Northeastern University

30. Rizvi, Noreen F. Genetic engineering of Catharanthus roseus to enhance production of anticancer pharmaceutical compounds.

Degree: PhD, Department of Chemical Engineering, 2015, Northeastern University

URL: http://hdl.handle.net/2047/D20194128

► The medicinal plant, Catharanthus roseus, is the source of pharmaceutically valuable anticancer alkaloids, vincristine (VCR) and vinblastine (VBL). These alkaloids are produced only in C.… (more)

Subjects/Keywords: metabolic engineering; RNAi; terpenoid indole alkaloids; transcription factor; Transgenic plants; Catharanthus roseus; Genetic engineering; Indole alkaloids; Synthesis; Terpenes; Transcription factors; RNA interference

Record Details Similar Records Cite « Share

❌

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

APA (6^th Edition):

Rizvi, N. F. (2015). Genetic engineering of Catharanthus roseus to enhance production of anticancer pharmaceutical compounds. (Doctoral Dissertation). Northeastern University. Retrieved from http://hdl.handle.net/2047/D20194128

Chicago Manual of Style (16^th Edition):

Rizvi, Noreen F. “Genetic engineering of Catharanthus roseus to enhance production of anticancer pharmaceutical compounds.” 2015. Doctoral Dissertation, Northeastern University. Accessed January 18, 2021. http://hdl.handle.net/2047/D20194128.

MLA Handbook (7^th Edition):

Rizvi, Noreen F. “Genetic engineering of Catharanthus roseus to enhance production of anticancer pharmaceutical compounds.” 2015. Web. 18 Jan 2021.

Vancouver:

Rizvi NF. Genetic engineering of Catharanthus roseus to enhance production of anticancer pharmaceutical compounds. [Internet] [Doctoral dissertation]. Northeastern University; 2015. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/2047/D20194128.

Council of Science Editors:

Rizvi NF. Genetic engineering of Catharanthus roseus to enhance production of anticancer pharmaceutical compounds. [Doctoral Dissertation]. Northeastern University; 2015. Available from: http://hdl.handle.net/2047/D20194128

Open Access Theses and Dissertations