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Targeted Gene Knock-out Via Promoter Tagging in Tomato (Lycopersicon Esculentum) and Tobacco (Nicotiana Tabacum).
Title
Targeted Gene Knock-out Via Promoter Tagging in Tomato (Lycopersicon Esculentum) and Tobacco (Nicotiana Tabacum).
Creator
Kazy, Alia, Zhang, Xing-Hai, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description
Many different ways to create mutants have been established. This research demonstrates yet another variation of the promoter tagging technique that allows for a single step selection of the putative transgenic plants that have a mutation in constitutively expressed genes. While tomato transformants have not yet been convincingly confirmed, tobacco transformation resulted in seven transgenic lines showing resistance to high concentrations of kanamycin. Two transgenic lines were further...
Show moreMany different ways to create mutants have been established. This research demonstrates yet another variation of the promoter tagging technique that allows for a single step selection of the putative transgenic plants that have a mutation in constitutively expressed genes. While tomato transformants have not yet been convincingly confirmed, tobacco transformation resulted in seven transgenic lines showing resistance to high concentrations of kanamycin. Two transgenic lines were further investigated and three putative promoters isolated. Transient expression analysis of leaves transformed by particle bombardment with vectors carrying beta-glucuronidase gene driven by these putative promoters suggests two of them to be functional. Further investigation is needed to confirm the expression in the stably transformed plants as well as cloning of the genes downstream of the functional promoters and research of their functions.
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Date Issued
2007
PURL
http://purl.flvc.org/fau/fd/FA00000779
Subject Headings
Gene mapping--Methodology, Gene mapping--Data processing, Gene expression, Microbial genetics, Computational biology
Format
Document (PDF)
Discovery and visualization of co-regulated genes relevant to target diseases
Title
Discovery and visualization of co-regulated genes relevant to target diseases.
Creator
Lad, Vaibhan., College of Engineering and Computer Science, Department of Computer and Electrical Engineering and Computer Science
Abstract/Description
In this thesis, we propose to discover co-regulated genes using microarray expression data, as well as providing visualization functionalities for domain experts to study relationships among discovered co-regulated genes. To discover co-regulated genes, we first use existing gene selection methods to select a small portion of genes which are relevant to the target diseases, on which we build an ordered similarity matrix by using nearest neighbor based similarity assessment criteria. We then...
Show moreIn this thesis, we propose to discover co-regulated genes using microarray expression data, as well as providing visualization functionalities for domain experts to study relationships among discovered co-regulated genes. To discover co-regulated genes, we first use existing gene selection methods to select a small portion of genes which are relevant to the target diseases, on which we build an ordered similarity matrix by using nearest neighbor based similarity assessment criteria. We then apply a threshold based clustering algorithm named Spectral Clustering to the matrix to obtain a number of clusters. The genes which are clustered together in one cluster represent a group of co-regulated genes and to visualize them, we use Java Swings as the tool and develop a visualization platform which provides functionalities for domain experts to study relationships between different groups of co-regulated genes; study internal structures within each group of genes, and investigate details of each individual gene and of course for gene function prediction. Results are analyzed based on microarray expression datasets collected from brain tumor, lung cancers and leukemia samples.
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Date Issued
2010
PURL
http://purl.flvc.org/FAU/2976447
Subject Headings
Genomics, Gene mapping, Cell transformation, Cellular signal transduction
Format
Document (PDF)
Hypoxia-regulated glial cell-specific gene therapy to treat retinal neovascularization
Title
Hypoxia-regulated glial cell-specific gene therapy to treat retinal neovascularization.
Creator
Biswal, Manas Ranjan., Charles E. Schmidt College of Science, Center for Complex Systems and Brain Sciences
Abstract/Description
Diabetic retinopathy is an ischemic retinal neovascular disease causing vision loss among adults. The studies presented involve the design and testing of a gene therapy vector to inhibit retinal revascularization, similar to that found in diabetic retinopathy. Gene therapy has proven to be an effective method to introduce therapeutic proteins to treat retinal diseases. Targeting a specific cell type and expression of therapeutic proteins according to the tissue microenvironment should have an...
Show moreDiabetic retinopathy is an ischemic retinal neovascular disease causing vision loss among adults. The studies presented involve the design and testing of a gene therapy vector to inhibit retinal revascularization, similar to that found in diabetic retinopathy. Gene therapy has proven to be an effective method to introduce therapeutic proteins to treat retinal diseases. Targeting a specific cell type and expression of therapeutic proteins according to the tissue microenvironment should have an advantage over traditional gene therapy by avoiding unwanted transgene expression. Hypoxia plays a significant role in the pathophysiology of many retinal ischemic diseases. Retinal Mèuller cells provide structural and functional support to retinal neurons, as well as playing a significant role in retinal neovascularization. Targeting Mèuller cells may be an effective strategy to prevent retinal neovascularization under pathological conditions. ... The hypoxia regulated, glial specific vector successfully reduced the abnormal neovascularization in the periphery by 93% and reduced the central vasobliterated area by 90%. A substantial amount of exogenous endostatin was produced in the retinas of P17 OIR mice. A significant increase in human endostatin protein and reduced vascular endothelial growth factor (VEGF) were identified by Western blot and ELISA, respectively. These findings suggest hypoxia-regulated, glial cell-specific scAAV mediated gene expression may be useful to prevent blindness found in devastating retinal diseases involving neovascularization.
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Date Issued
2012
PURL
http://purl.flvc.org/FAU/3359290
Subject Headings
Diabetic retinopathy, Research, Methodology, Gene therapy, Retinal degeneration, Treatment, Neovascularization inhibitors, Mitochondrial pathology, Retina, Cytology, Gene mapping
Format
Document (PDF)
Spatial analysis and functional gene clustering between lens epithelium and fiber cells
Title
Spatial analysis and functional gene clustering between lens epithelium and fiber cells.
Creator
Cowell, Tracy L., Florida Atlantic University, Kantorow, Marc
Abstract/Description
Purpose. To identify genes important for maintaining the specialized functions of lens epithelial and fiber cells. Methods. The expression profiles of 22,215 genes between human lens epithelial and fiber cells were analyzed using oligonucleotide microarray hybridization and RT-PCR. Selected genes were functionally clustered using the EASE bioinformatics software package. Results. Analysis of microarray hybridizations revealed 1430 transcripts that were significantly increased and 901...
Show morePurpose. To identify genes important for maintaining the specialized functions of lens epithelial and fiber cells. Methods. The expression profiles of 22,215 genes between human lens epithelial and fiber cells were analyzed using oligonucleotide microarray hybridization and RT-PCR. Selected genes were functionally clustered using the EASE bioinformatics software package. Results. Analysis of microarray hybridizations revealed 1430 transcripts that were significantly increased and 901 transcripts that were significantly decreased. Microarray data was confirmed using RT-PCR on 11 randomly selected genes. Functional clustering of the identified gene expression patterns revealed altered gene expression in cellular pathways including oxidative stress, cell proliferation, and apoptosis. The methionine sulfoxide reductase class of enzymes were further analyzed and demonstrated to be expressed throughout the human body, indicating a significant protective role. Conclusions. These data reveal novel and previously identified gene expression differences that provide insight into those mechanisms that may be important for lens cell differentiation.
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Date Issued
2004
PURL
http://purl.flvc.org/fcla/dt/13193
Subject Headings
Gene mapping--Statistical methods, Eye--Physiology, Epithelium--Culture and culture media, Cell culture--Analysis
Format
Document (PDF)