TRiP resources for modeling human disease

用于人类疾病建模的 TRiP 资源

• 批准号: 10598494
• 负责人: NORBERT PERRIMON
• 金额: $ 63.56万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10598494
• 关键词: Address; Affect; Animals; Area; Bioinformatics; Brain; CRISPR-mediated transcriptional activation; CRISPR/Cas technology; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Collection; Communities; Country; Data; Databases; Deposition; Disease; Drosophila genus; Funding; Funding Opportunities; Gene Activation; Gene Expression; Genes; Genetic; Genome; Genome engineering; Goals; Heart; Hereditary Disease; Home; Human; Individual; Kidney; Knock-in; Knock-out; Knowledge; Malignant Neoplasms; Mediating; Mission; Neurodegenerative Disorders; Organ; Orthologous Gene; Paper; Phenotype; Policies; Production; Proteins; Publications; R24; RNA Interference; Reagent; Repression; Request for Applications; Research; Research Design; Research Personnel; Research Project Grants; Resources; System; Technology; Time; Tissues; Transgenic Organisms; US State; United States National Institutes of Health; animal model development; combinatorial; fly; functional genomics; gain of function; gene conservation; gene function; genetic manipulation; genome-wide; genomic platform; human disease; human model; in vivo; knock-down; loss of function; medical schools; overexpression; response; small hairpin RNA; therapeutic target; tool

PROJECT SUMMARY / ABSTRACT This application requests continued funding for the TRiP, a highly successful in vivo Drosophila functional genomics platform at Harvard Medical School. To date we have generated more than 17,000 RNAi and CRISPR fly stocks for the research community. We propose to expand the TRiP resource of versatile and transformative transgenic tools for gene activation, repression and genome engineering. The goals of this resource, focused on the 3017 fly genes orthologous to genes known or suspected to be associated with human diseases, are to: (1) generate shRNA fly stocks for RNAi that address remaining gaps in the collection, such as genes not yet covered or reagents identified by the community as ineffective; (2) generate sgRNA stocks for CRISPR knock out (CRISPRko) and CRISPR activation (CRISPRa); (3) Build a toolkit of reagents that combine GAL4/UAS and a second binary expression system (LexA/lexAop or QF/QUAS) to facilitate RNAi and/or CRISPR control of gene expression in two different tissues independently; and (5) evaluate whole animal as well as tissue-specific loss-of-function (LOF) and gain-of-function (GOF) phenotypes associated with the new lines and curate information on the quality of individual RNAi and sgRNA lines in our “RSVP Plus” phenotype database. The result of these efforts will be a tremendous resource of fly stocks, distributed to the community by the Bloomington Drosophila Stock Center, which will allow researchers to easily knock down, knock out, or activate genes covered by the collection. When combined with the wide array of GAL4 lines, and our proposed new collection of LexA/QF lines, this genetic toolkit will allow researchers to modulate gene expression in any given stage and in multiple tissues simultaneously.

项目概要/摘要 该申请要求继续为 TRiP 提供资金，TRiP 是一种非常成功的果蝇体内功能 哈佛医学院的基因组学平台。迄今为止，我们已经生成了超过 17,000 个 RNAi 供研究界使用的 CRISPR 蝇种。我们建议扩大 TRiP 资源的多功能性和 用于基因激活、抑制和基因组工程的转化转基因工具。本次活动的目标 资源，重点关注 3017 个果蝇基因，这些基因与已知或疑似相关的基因同源 人类疾病的目标是：(1) 生成用于 RNAi 的 shRNA 果蝇种群，以解决收集中剩余的空白， 例如尚未涵盖的基因或被社区认定为无效的试剂； (2) 生成sgRNA 用于 CRISPR 敲除 (CRISPRko) 和 CRISPR 激活 (CRISPRa) 的库存； (3) 构建试剂工具包 结合 GAL4/UAS 和第二个二元表达系统（LexA/lexAop 或 QF/QUAS）以促进 RNAi 和/或 CRISPR 独立控制两个不同组织中的基因表达； (5) 评估整体 动物以及组织特异性功能丧失（LOF）和功能获得（GOF）表型与 新品系并在我们的“RSVP Plus”中整理有关单个 RNAi 和 sgRNA 品系质量的信息 表型数据库。这些努力的结果将是巨大的苍蝇资源，分发给 布卢明顿果蝇库存中心社区，这将使研究人员能够轻松击倒， 敲除或激活该集合所涵盖的基因。当与广泛的 GAL4 系列结合使用时， 我们提出的 LexA/QF 系新集合，该遗传工具包将允许研究人员调节基因 在任何给定阶段和多个组织中同时表达。