Genetics of long non-coding RNAs in zebrafish

斑马鱼长非编码RNA的遗传学

• 批准号: 8736103
• 负责人: ALEXANDER F SCHIER
• 金额: $ 35.07万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 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/8736103
• 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; Functional RNA; Gene Expression; Generations; Genes; Genetic; Genetic Screening; Genome; Genome Components; Genomics; Goals; Image; Knock-out; Larva; Length; Life; Location; Measurement; Mediating; Motor Activity; Mus; Mutation; Nucleotides; Organogenesis; Phenotype; Process; Proteins; RNA Splicing; Reagent; Research; Resources; Ribosomes; Role; Site; Structure; Technology; Transcript; Vertebrates; Zebrafish; base; endonuclease; genetic analysis; genome wide association study; in vivo; insight; mutant; novel; positional cloning; public health relevance; transcriptome sequencing; zebrafish development

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）：拟议研究的长期目标是确定长非编码RNA在脊椎动物发育中的功能。在过去的十年里，全基因组研究激增，这些研究鉴定出了大量以前未注释的转录本。在这些新的转录本中，长非编码 RNA (lncRNA) 类别引起了特别关注，因为它们代表了基因组中广泛的、很大程度上未经探索的组成部分：基因组的“暗物质”。拟议的研究旨在进行反向遗传筛选并分析斑马鱼中 100 个 lncRNA 的功能。基因组学和计算方法已经鉴定了斑马鱼发育过程中表达的数百个 lncRNA。根据保守性、基因组位置、表达和染色质谱选择的 100 个 lncRNA 将被破坏。反义寡核苷酸将用于直接干扰lncRNA，位点特异性核酸内切酶（TALEN和CRISPR）将用于破坏lncRNA基因。然后胚胎和幼虫将进行系统的表型表征，以揭示 lncRNA 的功能。拟议的反向遗传筛选将首次对脊椎动物 lncRNA 体内功能进行全面分析，并为该领域揭示这类神秘分子的作用提供资源。