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The Role of Alternative Splicing Factor Sfrs10 in Neural Development

选择性剪接因子 Sfrs10 在神经发育中的作用

基本信息

批准号：
8223205
负责人：
RAHUL N KANADIA
金额：
$ 20.53万
依托单位：
UNIVERSITY OF CONNECTICUT STORRS
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8223205
关键词：
Address Affect Alternative Splicing Amacrine Cells Antibodies Architecture Bioinformatics Biological Assay Biological Models Birth Order Brain Cell Separation Cells Central Nervous System Part Code Complementary DNA Complex Connecticut Data Defect Development Disease Electroporation Embryo Event Exclusion Exons Fluorescence Genes Glass Goals Grant House mice Human Immunofluorescence Immunologic In Situ Hybridization Infection Instruction Knockout Mice Laboratories Lead Left Link Messenger RNA Methods Microarray Analysis Motor Neurons Mus Mutation Myotonic Dystrophy Needles Neuraxis Neuroglia Neurons Nonsense Codon Northern Blotting Pathogenesis Pathway interactions Pattern Phenotype Plasmids Play Positioning Attribute Process Production Protein Isoforms Proteins Proteome RNA RNA Interference RNA Splicing Reporting Research Resolution Retina Retinal Retinitis Pigmentosa Reverse Transcriptase Polymerase Chain Reaction Role Science Spinal Muscular Atrophy Stem cells Techniques Tissues Tretinoin Universities Vertebrates Work base career cell type design gain of function ganglion cell glutamate receptor type B human fetus tissue imprint in vivo indexing injured insight nervous system development neurodevelopment postnatal professor relating to nervous system research study retinal damage retinal progenitor cell

项目摘要

Alternative .splicing allows hutTians to protiuce a vast proteome diversity from a relatively few (~24,000) protein coding genes. This ROO proposal is part: of an overall objective to gain insight into the fole of alternative splicing in retinal development. 1 have chosen to address this question by focusing on SfrslO, with the specific hypothesis that it is essential for cell fate determination and differentiation. The overall goal is to determine the expression, function and targets of SfrslO during retinal development. I have employed in situ hybridization, immunofluorescence and microarray analysis to determine the expression of SfrslO during retinal development. In all, SfrslO appears to be expressed in progenitor cells and differentiating amacrine cells. Interestingly, the microarray data obtained from single retinal cells shows that certain progenitor cell markers such as cycling Dl and Sfrsp2 are enriched along with the expression of SfrslO. To determine the function of SfrslO,! have employed in vivo electroporation to deliver F^NAi or the misexpression construct in PO retinal progenitor cells. 1 have found that loss of SfrslO function results in increase in the numberof Mullerglia at the expense of neurons. This experiment utilizes a Hamilton needle to deliver the plasmid into the subretinal space. However, this method has often created damage to the retina. 1 have now employed a different method which utilized glass needles that leave a negligible imprint on the retina. While RNAi constructs reported in the grant have been used to determine the function of SfrslO, a conditional knockout mouse is crucial to this proposal. The production of such a mouse has been initiated at the University of Connecticut, where I will begin my position as an assistant professor on August I'S 2010. As 1 continue this project in my laboratory at the University of Connecticut, I will further refine the expression of SfrslO, perform gain and loss of SfrslO function with the electroporation techinque. Finally, the conditional knockout mouse will be crossed to retina specific Cre lines followed by detailed analysis of the phenotype. Also, the knockout mouse retinal cells will be used for deep (454) sequecing to identify the targets of SfrslO. Extenstion of this work in my laboratory will provide me with preliminary data that 1 intend to use for an ROl grant that should launch my independent career in science.

的选择。剪接使赫特人能够从相对较少的（~ 24000）种蛋白质中产生大量的蛋白质组多样性