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Motor neuron selector genes and mechanism of their action

运动神经元选择基因及其作用机制

基本信息

批准号：
8890896
负责人：
Hynek Wichterle
金额：
$ 54.02万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8890896
关键词：
Address Adopted Algorithms Axon Binding Biochemical Biological Assay Cell physiology Cells ChIP-seq Chromatin Chromatin Structure Clinical Competence Computer Analysis DNA DNA Binding DNA-Protein Interaction Development Developmental Process Disease model Enhancers Event Failure Gene Expression Generations Genes Genetic Transcription Genomic Segment Genomics Individual Maps Methods Molecular Molecular Conformation Motor Neuron Disease Motor Neurons Muscle Mutation Analysis Ontology Output Pattern Pluripotent Stem Cells Population Process Recruitment Activity Refractory Regulation Reporter Signal Transduction Source Specificity Spinal Cord Staging System Testing Transplantation Universities cell motility cell type cholinergic clinical application clinically relevant design drug discovery embryonic stem cell genome-wide in vitro Assay in vivo insight nerve stem cell network models novel phenomenological models programs stem cell biology synaptogenesis syntax transcription factor

项目摘要

DESCRIPTION (provided by applicant): Transcriptional programming of cell identity is gaining importance at both basic developmental and clinical level. While the phenomenology of cell programming and reprogramming by forced expression of transcription factors is well described, the mechanism of action of programming factors or the sequence of regulatory events resulting in a cell adopting a new identity are largely unknown. We propose to combine the strengths of stem cell biology with genomic and computational approaches to characterize the process of transcriptional programming of motor neuron (MN) identity. We developed efficient methods for the induction of MN identity in differentiating embryonic stem cells (ESCs) by the expression of defined transcription factors. Using the system we will combine biochemical, genomic and computational analysis to address following questions: i) whether programming factors directly regulate terminal motor neuron effector genes or initiate a cascade of intermediate transcription programs; ii) whether recruitment of programming factors to DNA is cooperative and which factors determine the specificity of DNA binding; iii) whether identified MN enhancers are inaccessible for programming factor binding in cell types refractory to MN programming; iv) whether identified secondary binding motifs for Onecut and Ebf transcription factors contribute to productive regulation of MN specific gene expression; v) whether additional secondary motifs recruit ancillary transcription factors to NIL bound enhancers that contribute to productive regulation of MN specific gene expression. Together these studies will provide fundamental insight into the developmental processes underlying specification of defined cell identity and will provide novel and efficient source of motor neurons for disease modeling, study and drug discovery.

描述（由申请人提供）：细胞身份的转录编程在基础发育和临床水平上都越来越重要。虽然通过转录因子的强制表达对细胞编程和重编程的现象进行了很好的描述，但编程因子的作用机制或导致细胞采用新身份的调控事件序列在很大程度上是未知的。我们建议将干细胞生物学的优势与基因组学和计算方法相结合，以表征运动神经元（MN）身份的转录编程过程。我们开发了有效的方法来诱导分化胚胎干细胞（ESCs）中MN的身份，通过表达定义的转录因子。使用该系统，我们将结合生化，基因组学和计算分析来解决以下问题：i)编程因子是否直接调节终端运动神经元效应基因或启动一系列中间转录程序；ii)编程因子对DNA的招募是否合作，哪些因素决定了DNA结合的特异性；iii)在对MN编程难的细胞类型中，已鉴定的MN增强子是否无法与编程因子结合；iv) oneccut和Ebf转录因子的二级结合基序是否有助于MN特异性基因表达的生产性调控；v)额外的次级基序是否将辅助转录因子招募到NIL结合增强子中，从而有助于MN特异性基因表达的生产性调节。总之，这些研究将为定义细胞身份和意志的发育过程提供基本的见解