TRiP resources for modeling human disease

用于人类疾病建模的 TRiP 资源

基本信息

批准号：
10374128
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
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10374128
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 Geographic state Goals Heart Hereditary Disease Human Individual Institutes Kidney Knock-in Knock-out Knowledge Malignant Neoplasms Mediating Mission Monoclonal Antibody R24 Neurodegenerative Disorders Organ Orthologous Gene Paper Phenotype Policies Production Proteins Publications RNA Interference Reagent Repression Request for Applications Research Research Design Research Personnel Research Project Grants Resources System Technology Time Tissues Transgenic Organisms United States National Institutes of Health animal model development base combinatorial fly functional genomics gain of function 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

项目摘要

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迄今为止，我们已经在哈佛医学院的基因组学平台上生成了超过 17,000 个 RNAi 和我们建议为研究界扩展 TRiP 资源。用于基因激活、抑制和基因组工程的转化转基因工具。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鉴定为无效的试剂（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；和 (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 品系质量的信息这些努力的结果将是一个巨大的蝇种群资源，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，当与广泛的 GAL4 系组合时，敲除或激活该集合所涵盖的基因，并且我们提议的 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。