Regulating transcription of the key neural lineage driver ASCL1

调节关键神经谱系驱动 ASCL1 的转录

基本信息

批准号：
10550431
负责人：
Jane E Johnson
金额：
$ 7.72万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10550431
关键词：
3-Dimensional ASCL1 gene ATAC-seq Autonomic nervous system Biological Assay Biological Models Brain CRISPR interference CRISPR/Cas technology Cell Culture Techniques Cells Chromatin Complex Data Development Developmental Gene Disease Distal Fibroblasts Future Gene Expression Gene Expression Profile Generations Genes Genetic Transcription Genome Genomic Segment Genomics Goals Human Knowledge Maps Modeling Molecular Molecular Conformation Mus Neural Tube Development Neurodegenerative Disorders Neuronal Differentiation Neurons Nuclear Pore Complex Olfactory Epithelium Population Process Protocols documentation Regulation Regulatory Element Reporter Retina Signal Transduction Site Spinal Cord Technology Testing Tissues Transcriptional Regulation Transgenic Mice Untranslated RNA cell type design experimental study human model in vitro Model in vivo in vivo Model in vivo evaluation induced pluripotent stem cell insight interest mouse genome mouse model nerve stem cell neurodevelopment neurogenesis neuropsychiatric disorder novel therapeutics postnatal progenitor relating to nervous system sensory system stem cells success transcription factor

项目摘要

The bHLH transcription factor ASCL1 (HASH1/MASH1) is essential for neuronal differentiation and sub-type specification of multiple neuronal cell-types throughout the brain, spinal cord, and autonomic nervous system, as well as cells in sensory systems such as the retina and olfactory epithelia. ASCL1 function is balanced with NOTCH signaling activity to control progenitor proliferation and differentiation. ASCL1 has also been identified as a pioneering factor and a key component of cocktails directly reprogramming fibroblasts to neurons. With these important functions attributed to ASCL1, and its requirement for controlled spatial and temporal expression in vivo for viability postnatally, it is surprising how little is known about regulation of ASCL1 gene transcription. This gap in knowledge reflects past technical challenges in identifying and manipulating cis-regulatory elements (REs) found at large distances from the gene of interest. REs functioning at long-distances to control key developmental genes are being discovered using advances in technologies that can interrogate and manipulate the spatial genome. Here we will exploit these technologies to gain much needed insights into transcriptional control of ASCL1 using cell culture and in vivo models of neural development. Each model has a particular strength that allows unique aspects of ASCL1 regulation to be uncovered. Aims include identifying and testing functions of long-range REs controlling ASCL1 during neuronal differentiation in mouse (in vivo) and human (in vitro) models. Success in these aims will provide functional non-coding regulatory sequences controlling ASCL1 expression. This is important for future projects to identify molecular components of the signaling complexes working through these REs to reach the goal of providing an understanding of how a key lineage defining transcriptional regulator is controlled during development and disease.

BHLH转录因子ASCL1（HASH1/MASH1）对于神经元分化至关重要以及整个大脑，脊髓和自主神经系统以及视网膜和嗅觉等感觉系统中的细胞上皮。 ASCL1功能与Notch信号活性平衡以控制祖细胞增殖和分化。 ASCL1也已被确定为开创性因素和关键因素鸡尾酒的成分直接将成纤维细胞重新编程为神经元。这些很重要归因于ASCL1的功能及其对受控空间和时间的要求在产后生存能力的体内表达，令人惊讶的是，对调节的调节知之甚少 ASCL1基因转录。知识的差距反映了过去的技术挑战识别和操纵顺式调节元件（RES）在距离感兴趣的基因。在控制关键发育基因的长期功能的RES功能是使用可以审问和操纵的技术的进步发现空间基因组。在这里，我们将利用这些技术来获得急需的见解使用细胞培养和神经发育的体内模型对ASCL1的转录控制。每个模型都有一个特殊的强度，允许ASCL1调节的独特方面为裸露。目的包括识别和测试远程RES控制ASCL1的功能在小鼠（体内）和人（体外）模型中的神经元分化过程中。这些成功 AIMS将提供控制ASCL1表达的功能非编码调节序列。这对于未来的项目识别信号复合物的分子成分很重要通过这些RES来实现对如何关键的理解的目标定义转录调节剂的谱系在发育和疾病期间受到控制。