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The effect of histone post-translational modification on transcriptional bursting during development

组蛋白翻译后修饰对发育过程转录爆发的影响

基本信息

批准号：
10401153
负责人：
Joseph M Zinski
金额：
$ 5.87万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10401153
关键词：
3&apos Untranslated Regions Acetylation Acylation Affect Binding Biological Assay CRISPR imaging Cell Cycle Cells ChIP-seq Characteristics Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Complex DNA DNA-Directed RNA Polymerase Data Development Disease Drosophila genus EP300 gene Embryo Embryonic Development Enhancers Equilibrium Etiology Eukaryota Exhibits Frequencies Gene Expression Genes Genetic Transcription Goals Heterogeneity Histone Deacetylase Histones Image Individual Kinetics Label Length Link Malignant Neoplasms Measurement Measures Mediating Messenger RNA Methods Methylation Modeling Modification Molecular Oocytes Pattern Play Polycomb Post-Translational Protein Processing Prevalence Production RNA RNA Interference Regulation Reporter Role Site Stochastic Processes Testing Transcriptional Regulation Transgenes Work cell fate specification chromatin remodeling effective therapy efficacy testing experimental study fluorescence imaging genome-wide genomic locus histone acetyltransferase histone demethylase histone methyltransferase histone modification in vivo knock-down mathematical model nuclease promoter theories transcriptome sequencing

项目摘要

Title: The effect of histone post-translational modification on transcriptional bursting during development In all eukaryotes studied, transcription at individual gene loci exhibits random oscillations between active and inactive states, a phenomenon known as transcription bursting, which fundamentally impacts the synchrony and robustness of cell fate specification during embryonic development. To understand how transcription controls development, it is necessary to uncover the molecular determinants that control the duration of bursts and the rates at which bursts occur. Eukaryotic transcription is requires chromatin remodelers to change the chromatin state at enhancers and promoters. It is unclear what effect chromatin remodeling has on transcription burst lengths and frequencies. Here, I will knock down an array of chromatin remodelers, histone methyltransferases, and histone acetylases during Drosophila oocyte development and determined their effect on early axis patterning. For those that effect patterning, I will determine how transcriptional burst lengths and frequencies are modified in target genes. I will identify target genes using RNA-seq and CHIP-seq. To querry changes in transcriptional bursting, I will employ single mRNA FISH and live nascent site imaging. To quantitatively describe state transitions using these assays, I will use a 2-state mathematical model of transcription states. Together, these experiments show how changes in chromatin states change transcriptional bursting dynamics in target genes. I am requesting an additional nine months to complete my work from June 1st, 2021 through March 31st, 2022.

组蛋白翻译后修饰对发育过程中转录爆发的影响在所有研究的真核生物中，单个基因位点的转录表现出活性和活性之间的随机振荡。不活跃状态，一种被称为转录爆发的现象，它从根本上影响了和胚胎发育过程中细胞命运规范的稳健性。为了了解转录是如何控制发展，有必要揭示控制爆发持续时间的分子决定因素以及突发发生的速率。真核转录需要染色质重塑剂来改变细胞的结构。在增强子和启动子处的染色质状态。目前尚不清楚染色质重塑对细胞有何影响转录爆发长度和频率。在这里，我将敲除一系列的染色质重塑，组蛋白甲基转移酶和组蛋白乙酰化酶在果蝇卵母细胞发育过程中的作用关于早期轴模式对于那些影响模式的，我将确定转录爆发长度和在靶基因中改变频率。我将使用RNA-seq和CHIP-seq鉴定靶基因。去询问在转录爆发的变化中，我将采用单个mRNA FISH和活的新生位点成像。到定量描述状态转换使用这些分析，我将使用一个2状态的数学模型，转录状态。总之，这些实验显示了染色质状态的变化靶基因的转录爆发动力学。我请求再延长9个月来完成我的工作时间为2021年6月1日至2022年3月31日。