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Transcriptional and Post-transcriptional Regulation of Gene Expression
Title
Transcriptional and Post-transcriptional Regulation of Gene Expression.
Creator
Ding, Jun, Hu, Haiyan, Li, Xiaoman, Zhang, Shaojie, Jin, Yier, University of Central Florida
Abstract / Description
Regulation of gene expression includes a variety of mechanisms to increase or decrease specific gene products. Gene expression can be regulated at any stage from transcription to post-transcription and it's essential to almost all living organisms, as it increases the versatility and adaptability by allowing the cell to express the needed proteins. In this dissertation, we comprehensively studied the gene regulation from both transcriptional and post-transcriptional points of view....
Show moreRegulation of gene expression includes a variety of mechanisms to increase or decrease specific gene products. Gene expression can be regulated at any stage from transcription to post-transcription and it's essential to almost all living organisms, as it increases the versatility and adaptability by allowing the cell to express the needed proteins. In this dissertation, we comprehensively studied the gene regulation from both transcriptional and post-transcriptional points of view. Transcriptional regulation is by which cells regulate the transcription from DNA to RNA, thereby directing gene activity. Transcriptional factors (TFs) play a very important role in transcriptional regulation and they are proteins that bind to specific DNA sequences (regulatory elements) to regulate the gene expression. Current studies on TF binding are still very limited and thus, it leaves much to be improved on understanding the TF binding mechanism. To fill this gap, we proposed a variety of computational methods for predicting TF binding elements, which have been proved to be more efficient and accurate compared with other existing tools such as DREME and RSAT peaks-motif. On the other hand, studying only the transcriptional gene regulation is not enough for a comprehensive understanding. Therefore, we also studied the gene regulation at the post-transcriptional level. MicroRNAs (miRNAs) are believed to post-transcriptionally regulate the expression of thousands of target mRNAs, yet the miRNA binding mechanism is still not well understood. In this dissertation, we explored both the traditional and novel features of miRNA-binding and proposed several computational models for miRNA target prediction. The developed tools outperformed the traditional microRNA target prediction methods (.e.g miRanda and TargetScan) in terms of prediction accuracy (precision, recall) and time efficiency.
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Date Issued
2016
Identifier
CFE0006098, ucf:51197
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0006098
DETERMINANTS OF CHLOROPLAST GENE EXPRESSION AND APPLICATIONS OF CHLOROPLAST TRANSFORMATION IN LACTUCA SATIVA AND NICOTIANA TABACUM.
Title
DETERMINANTS OF CHLOROPLAST GENE EXPRESSION AND APPLICATIONS OF CHLOROPLAST TRANSFORMATION IN LACTUCA SATIVA AND NICOTIANA TABACUM.
Creator
Ruhlman, Tracey, Daniell, Henry, University of Central Florida
Abstract / Description
Genetic modification of plastids in the model plant tobacco (Nicotiana tabacum) has demonstrated that numerous foreign gene products can accumulate to high levels in this setting. Plastid biotechnology is maturing to encompass the improvement of food and feed species and the production of biopharmaceutical proteins for oral delivery necessitating development of stable transplastomic edible plants. In the interest of establishing an edible platform we have investigated the use of native and...
Show moreGenetic modification of plastids in the model plant tobacco (Nicotiana tabacum) has demonstrated that numerous foreign gene products can accumulate to high levels in this setting. Plastid biotechnology is maturing to encompass the improvement of food and feed species and the production of biopharmaceutical proteins for oral delivery necessitating development of stable transplastomic edible plants. In the interest of establishing an edible platform we have investigated the use of native and foreign regulatory elements in relation to foreign gene expression in plastids. Multiple sequence alignments of intergenic regions for 20 species of angiosperm showed that despite 95% identity in the coding region, identity in the psbA upstream region is 59% across all taxa examined, other gene coding regions displayed sequence identity of 80-97%, whereas the non-coding regions were 45-79% suggesting that our physical data can be extrapolated beyond the model presented. We found that by exchanging psbA untranslated regions (UTRs) between N. tabacum and lettuce (Lactuca sativa), the expression of the CTB-proinsulin (CTB-Pins) monocistronic transcript declined by 84% and foreign protein accumulation was reduced by as much as 97% in mature leaves. Polyribosome association assays suggest that ribosome-free transgenic transcripts are stabilized where the native UTR is employed. RNA EMSA revealed that binding proteins interacted with psbA 5' UTRs in a species specific manner and the half life of the L. sativa 5'UTR-CTB-Pins mRNA was reduced by 3.7 fold in N. tabacum stromal extracts. Our data indicate that the use of species-specific regulatory elements could lead to establishment of reproducible plastid transformation in desirable target species such as L. sativa. Using transplastomic L. sativa for oral delivery of bioencapsulated CTB-Pins we delayed the onset of diabetes in NOD mice when retinyl acetate supplement was provided compared to untouched mice. In this 30 week study we monitored blood glucose levels and evaluated the in vitro suppressive capacity of regulatory T cells isolated from diabetic mice. Whether delay or prevention was achieved appeared to be a function of antigen dose as high dose resulted in a nine week delay of onset while low dose reduced the incidence of diabetes by 36%. In addition we have evaluated metabolic engineering in the N. tabacum model where we generated cis-genic lines expressing nucleus-encoded methionine pathway enzymes in plastids. Transplastomic expression of Cystathionine gamma-Synthase led to a three-fold increase in enzyme activity and a doubling of methionine content in leaves without a deleterious phenotype. In exploring molecular mechanisms supporting gene expression in plastids and applying transplastomic technology to real human problems this work seeks address the potential of plastid biotechnology for improvement of commodity crops and production of biopharmaceuticals.
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Date Issued
2009
Identifier
CFE0002687, ucf:48236
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0002687