Genetics of long non-coding RNAs in zebrafish

斑马鱼长非编码RNA的遗传学

基本信息

批准号：
9056624
负责人：
ALEXANDER F SCHIER
金额：
$ 34.72万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-15 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9056624
关键词：
Address Antisense Oligonucleotides Attention Behavioral Bioinformatics Biological Models Biological Process Cellular biology Chromatin Chromatin Structure Code Complement Complex Data Development Embryo Embryonic Development Explosion Gene Expression Generations Genes Genetic Genetic Screening Genome Genome Components Genomics Goals Health Knock-out Larva Length Life Location Measurement Mediating Motor Activity Mus Mutation Nucleotides Organogenesis Phenotype Process Proteins RNA Splicing Reagent Research Resources Role Site Structure Technology Transcript Untranslated RNA Vertebrates Zebrafish base dark matter endonuclease genetic analysis genome-wide analysis imaging biomarker in vivo insight mutant novel reverse genetics ribosome profiling transcription activator-like effector nucleases transcriptome sequencing zebrafish development

项目摘要

DESCRIPTION (provided by applicant): The long-range goal of the proposed studies is to determine the functions of long non-coding RNAs in vertebrate development. The past decade has seen an explosion of genome-wide studies that have identified a plethora of previously unannotated transcripts. Among these novel transcripts, the class of long non-coding RNAs (lncRNAs) has attracted particular attention since they represent an extensive, largely unexplored component of the genome: the "dark matter" of the genome. The proposed study aims to perform a reverse genetic screen and analyze the function of 100 lncRNAs in zebrafish. Genomics and computational approaches have led to the identification of several hundred lncRNAs expressed during zebrafish development. 100 of these lncRNAs, selected by conservation, genomic location, expression, and chromatin profile, will be disrupted. Antisense oligonucleotides will be used to directly interfere with lncRNAs, and site-specific endonucleases (TALENs and CRISPRs) will be employed to disrupt lncRNA genes. Embryos and larvae will then undergo systematic phenotypic characterization to reveal the functions of lncRNAs. The proposed reverse genetic screen will provide the first comprehensive analysis of vertebrate lncRNA function in vivo and generate the resources for the field to reveal the roles of this mysterious class of molecules.

描述（由申请人提供）：拟议的研究的远程目标是确定长期非编码RNA在脊椎动物发育中的功能。在过去的十年中，全基因组研究的爆炸激增，已经确定了许多先前未经宣传的转录本。在这些新颖的转录本中，长期非编码RNA（LNCRNA）的类别引起了人们的特别关注，因为它们代表了基因组的广泛，很大程度上未开发的成分：基因组的“暗物质”。拟议的研究旨在执行反向遗传筛查，并分析斑马鱼中100个LNCRNA的功能。基因组学和计算方法导致鉴定了斑马鱼发育过程中表达的数百个lncRNA。这些LNCRNA中有100个是由保护，基因组位置，表达和染色质谱选择的。反义寡核苷酸将用于直接干扰LNCRNA，并且将使用特定位点的核酸酶（Talens and CRISPR）来破坏LNCRNA基因。然后，胚胎和幼虫将经历系统的表型表征，以揭示LNCRNA的功能。所提出的反向遗传筛查将对体内脊椎动物lncRNA功能进行首次综合分析，并为该领域生成资源以揭示这种神秘的分子类别的作用。