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

• 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
• 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' 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

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.

标题：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跃和活跃之间的随机振荡。 非活动状态，一种称为转录爆发的现象，从根本上影响同步 以及胚胎发育过程中细胞命运规范的稳健性。了解如何转录 控制发育，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者来改变 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有何影响 转录突发长度和频率。在这里，我将敲除一系列染色质重塑因子，组蛋白 果蝇卵母细胞发育过程中的甲基转移酶和组蛋白乙酰化酶并确定其作用 关于早期的轴图案。对于那些影响模式的人，我将确定转录突发长度和 频率在目标基因中被修改。我将使用 RNA-seq 和 CHIP-seq 来识别目标基因。查询 转录爆发的变化，我将采用单 mRNA FISH 和活新生位点成像。到 使用这些分析定量描述状态转换，我将使用 2 态数学模型 转录状态。这些实验共同展示了染色质状态的变化是如何变化的 靶基因的转录爆发动力学。我请求额外九个月的时间来完成我的工作 工作时间为2021年6月1日至2022年3月31日。