Mechanisms regulating Evf2 long non-coding RNA transcriptional control

Evf2长链非编码RNA转录控制的调控机制

基本信息

批准号：
8824967
负责人：
Jhumku Dutt Kohtz
金额：
$ 41.76万
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：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8824967
关键词：
Adult Alleles Autistic Disorder Biochemistry Biological Brain Caliber Cancer Control Cell Nucleus Chromatin Remodeling Factor Complex DNA DNA Methylation DNA Modification Methylases Development Dimensions Disease Dosage Compensation (Genetics)Embryo Enhancers Epilepsy Erinaceidae Family Family member Gene Dosage Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genome Genomic Segment 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 Untranslated RNA 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）：尽管已经建立了一段时间在开发调节的转录因子和分化中的关键作用，但长期非编码RNA（LNCRNA）的作用才开始被理解。我的实验室发现了新型的非编码RNA，EVF2和EVF1，这些RNA调节了适当的GABA能中间神经元开发至关重要的转录因子。我们在EVF2上的工作，一种被转录为剪接的反义转录本的LNCRNA表明LNCRNA具有特定的基因调节函数。 EVF2是跨脊椎动物（超级保守）的保守的，并且在发育中的前脑中被声波刺猬激活。我们表明，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。通过分析我实验室中开发的EVF2功能丧失小鼠模型，我们发现EVF2不仅需要适当的DLX5和DLX6转录调节，而且还需要适当的GABAergic Interneuron interneuron在胚胎脑和成人海马电路中的适当性GABAergic Interneuron发育。尽管我过去的大部分工作都集中在EVF2 lncRNA在间神经元开发中的生物学作用上，但EVF2长期非编码RNA依赖性转录调控所涉及的精确机制仍然未知。拟议的研究将使用小鼠遗传学，RNA和蛋白质生物化学以及胚胎大脑核中的RNA云可视化来确定EVF2 LNCRNA转录调控的机制。某些技术是新颖的，例如在三个维度中使用RNA云可视化，单位和双性分析以及体外 DNA依赖性RNA蛋白质复合物组件；整个方法是新颖的，因为尚未报道研究lncRNA基因调节方法的这种组合。这些研究将提供对基因调节的关键方面的理解，并与神经发育和功能以及生长控制和癌症特别相关，在这些研究中，已经确定了多个超保守的LNCRNA。