High-Throughput Analysis of Tissue/Cell-Type-Specific Cis Regulatory Modules

在后生动物中，基因表达是以组织/细胞类型特有的方式调节的 主要通过称为顺式调控的非编码序列的延伸 调节(通常)邻近基因(S)表达的模块(CRM)。CRM 包含一个或多个DNA结合位点，用于一个或多个序列特异性、调节性 转录因子(TF)激活或抑制靶基因表达 基因(S)；激活基因表达的CRM通常被称为转录 “增强剂”，而那些抑制基因表达的被称为“消音器”。 此更新项目的目标是扩展我们的‘Enhancer-FACS-Seq’(EFS) 用于分析附加功能的高通量实验分析技术 果蝇全胚胎体内组织/细胞类型特异性CRM的特征 基因组背景，并利用它们来识别控制基因的顺式调控密码 以特定于组织和细胞类型的方式表达。我们将专注于发展 以果蝇胚胎体细胞中胚层为模型系统。我们预计我们的 方法和数据应适用于其他系统和生物体，包括 人类。

In metazoans, gene expression is regulated in a tissue/cell-type specific manner predominantly via stretches of noncoding sequence referred to as cis regulatory modules (CRMs) that regulate the expression of (typically) the adjacent gene(s). CRMs contain 1 or more DNA binding sites for 1 or more sequence-specific, regulatory transcription factors (TFs) that function to activate or repress the expression of target gene(s); CRMs that activate gene expression are frequently referred to as transcriptional “enhancers”, while those that repress gene expression are referred to as “silencers”. The goals of this renewal project are to extend our ‘enhancer-FACS-Seq’ (eFS) technologies for high-throughput experimental assays to analyze additional functional features of tissue/cell-type-specific CRMs in vivo in whole Drosophila embryos in a genomic context, and to use them to identify cis regulatory codes that control gene expression in a tissue- and cell-type-specific manner. We will focus on the developing embryonic somatic mesoderm in Drosophila as our model system. We anticipate that our approaches and data should be applicable to other systems and organisms, including human.