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Elucidating an Xist-dependent program of sexually dimorphic alternative splicing in the mammalian brain

阐明哺乳动物大脑中依赖于 Xist 的性二态选择性剪接程序

基本信息

批准号：
9922380
负责人：
Douglas L Black
金额：
$ 64.17万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9922380
关键词：
Affect Alleles Alternative Splicing Automobile Driving Binding Binding Sites Biological Assay Brain Cells Computing Methodologies Data Data Set Databases Dependence Development Disease Event Excision Exhibits Female Four Core Genotypes Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Determinism Genetic Transcription Genotype Genotype-Tissue Expression Project Gonadal Steroid Hormones Hormone use Human Immunoprecipitation Knock-out Measurement Measures Mental disorders Messenger RNA Methods Molecular Mus Neuronal Differentiation Neurons Pattern Polypyrimidine Tract-Binding Protein Post-Transcriptional Regulation Predisposition Proteins RNA RNA Splicing RNA-Binding Proteins Regulation Reverse Transcriptase Polymerase Chain Reaction Role Source Structure Tissue-Specific Gene Expression Tissues Transcript Transgenic Mice Transgenic Organisms Untranslated RNA Woman X Chromosome X Inactivation base brain tissue crosslink dimorphism gene function genome-wide genomic locus mRNA Precursor male men multiple datasets mutant nervous system disorder neuropsychiatry prediction algorithm programs relating to nervous system sexual dimorphism tool transcriptome transcriptome sequencing

项目摘要

PROJECT SUMMARY Women and men are well known to have different propensities to neuropsychiatric illness, but the source of these differences is not understood. In particular, the molecular events that determine functional dimorphism between the female and male brain need to be defined. We will examine how the female-specific long noncoding RNA Xist and its newly identified interaction with the pre-mRNA splicing regulators PTBP1 and 2 affect gene expression and alternative splicing in the female brain. The project will use expertise and tools developed in three labs for the study of Xist RNA, of neuronal splicing regulation by the PTB proteins, and of gene expression and alternative splicing using computational methods. RNA-seq data from defined regions of both human and mouse brain will be analyzed using the new rMATS analysis tool to create a large statistically robust database of differential gene expression and alternative pre-mRNA splicing between males and females. Expression and splicing changes will be correlated with changes in PTBP1/2 mRNA and Xist across the same datasets to define genes potentially regulated by these molecules at the transcriptional and post- transcriptional levels. Female specific patterns of expression and splicing caused by the XX genotype will be distinguished from events driven by female hormones using four core genotype mice. Xist targeting will be confirmed using conditional Xist alleles that allow either removal or activation of Xist during brain development and measurement of the resulting changes in splicing. The expression of Xist relative to PTBP2 will be quantified over neuronal differentiation in culture. The PTBP targeting of Xist-dependent changes in splicing will be confirmed in PTBP2 knockout and PTBP1 transgenic mice, and by transcriptome-wide binding analyses by iCLIP. Alternative splicing is a widespread mechanism of gene regulation, but has been only minimally examined in relation to the XX genotype of female cells. Using sophisticated new genome-wide methods and molecular tools, these studies promise to identify new genetic determinants of sexual dimorphism in the mammalian brain and to elucidate their underlying molecular mechanisms. In the longer term, the identified molecular changes driven by Xist and PTBP will provide entrée to the examination of the functional consequences of these dimorphisms.

项目总结