Mechanisms regulating Evf2 long non-coding RNA transcriptional control

Evf2长非编码RNA转录控制的调控机制

基本信息

批准号：
8662396
负责人：
Jhumku Dutt Kohtz
金额：
$ 5.55万
依托单位：
LURIE CHILDREN'S HOSPITAL OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-10 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8662396
关键词：
Adult Alleles Autistic Disorder Biochemistry Biological Brain Caliber Cancer Control Cell Nucleus Chromatin Remodeling Factor Complex DNA DNA Methylation DNA Methyltransferase DNA Modification Methylases Development Dimensions Disease Dosage Compensation (Genetics)Embryo Enhancers Epilepsy Erinaceidae Family Family member Functional RNA Gene Dosage Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genome Genomics Germ Cells Grant Growth Heterogeneity Hippocampus (Brain)Human Imagery In Vitro Intergenic DNA Interneuron function Interneurons Laboratory Finding Lead Link Malignant Neoplasms Mass Spectrum Analysis Mediating Mental Retardation Methods Methylation Molecular Molecular Biology Mono-S Mus Nature Neurons Neurophysiology - biologic function Neurosciences Nuclear Nuclear Extract Nuclear RNA Nucleic Acid Regulatory Sequences Pattern Play Poly A Prosencephalon Protein Biochemistry Publishing RNA RNA Biochemistry RNA Splicing Recruitment Activity Regulator Genes Regulatory Element Reporting Role SMARCA4 gene Schizophrenia Techniques Time Transcript Transcription Coactivator Transcriptional Regulation Translating Vertebrates Work X Inactivation base cell growth homeodomain imprint loss of function malignant neurologic neoplasms member mouse model mutant nanometer nerve stem cell nervous system disorder neurodevelopment neuropsychiatry news novel parental influence protein complex research study structural biology subventricular zone transcription factor

项目摘要

DESCRIPTION (provided by applicant): Although a critical role for developmentally regulated transcription factors in patterning and differentiation has been established for some time, the role of long non-coding RNAs (lncRNAs) is only beginning to be understood. My lab discovered novel non-coding RNAs, Evf2 and Evf1, that regulate transcription factors essential for proper GABAergic interneuron development. Our work on Evf2, an lncRNA transcribed as a spliced anti-sense transcript shows that lncRNAs play specific gene regulatory functions. Evf2 is conserved across vertebrates (ultraconserved), and activated by sonic hedgehog in the developing forebrain. We show that Evf2 forms a complex with the DLX2 homeodomain transcription factor and regulates Dlx5/6 enhancer activity. Through analysis of an Evf2 loss-of-function mouse model developed in my lab, we find that Evf2 is not only required for proper transcriptional regulation of Dlx5 and Dlx6, but also for proper GABAergic interneuron development in embryonic brain and adult hippocampal circuitry . While much of my past work has focused on the biological role of Evf2 lncRNA in interneuron development, the precise mechanisms involved in Evf2 long- non-coding RNA-dependent transcriptional regulation remain unknown. The proposed studies will use mouse genetics, RNA and protein biochemistry, and RNA cloud visualization in nuclei of embryonic brain to determine the mechanism of Evf2 lncRNA transcriptional regulation. Some of the techniques are novel, such as the use of RNA cloud visualization in 3 dimensions, mono and bi-allelic analysis, and in vitro DNA-dependent RNA-protein complex assembly; the approach as a whole is novel, as this combination of methods to study lncRNA gene regulation has not been reported. These studies will provide an understanding of a critical aspect of gene regulation with particular relevance to neural development and function, as well as growth control and cancer, where multiple ultraconserved lncRNAs have been identified.

描述（由申请人提供）：虽然发育调节转录因子在模式形成和分化中的关键作用已经确定了一段时间，但长非编码 RNA (lncRNA) 的作用才刚刚开始被理解。我的实验室发现了新型非编码 RNA Evf2 和 Evf1，它们调节 GABA 能中间神经元正常发育所必需的转录因子。我们对 Evf2（一种转录为剪接反义转录本的 lncRNA）的研究表明，lncRNA 发挥特定的基因调控功能。 Evf2 在脊椎动物中是保守的（超保守的），并由发育中的前脑中的音刺猬激活。我们发现 Evf2 与 DLX2 同源域转录因子形成复合物并调节 Dlx5/6 增强子活性。通过对我实验室开发的 Evf2 功能丧失小鼠模型的分析，我们发现 Evf2 不仅是 Dlx5 和 Dlx6 正确转录调节所必需的，而且也是胚胎大脑和成年海马回路中 GABA 能中间神经元正常发育所必需的。虽然我过去的大部分工作都集中在 Evf2 lncRNA 在中间神经元发育中的生物学作用，但 Evf2 长链非编码 RNA 依赖性转录调节所涉及的精确机制仍然未知。拟议的研究将利用小鼠遗传学、RNA和蛋白质生物化学以及胚胎脑细胞核中的RNA云可视化来确定Evf2 lncRNA转录调控的机制。其中一些技术是新颖的，例如使用 3 维 RNA 云可视化、单等位基因和双等位基因分析以及体外 DNA依赖性RNA-蛋白质复合物组装；该方法整体上是新颖的，因为这种研究 lncRNA 基因调控的方法组合尚未见报道。这些研究将提供对基因调控的一个关键方面的理解，特别是与神经发育和功能以及生长控制和癌症相关的基因调控，其中已鉴定出多种超保守的lncRNA。