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：
10206288
负责人：
NORBERT PERRIMON
金额：
$ 63.37万
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/10206288
关键词：
Address Affect Animals Area Bioinformatics Brain 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.

项目摘要 /摘要该申请要求继续为这次旅行提供资金，这是一个非常成功的果蝇功能哈佛医学院的基因组学平台。迄今为止，我们已经产生了17,000多个RNAi和 CRISPR飞行研究界。我们建议扩大多功能和用于基因激活，反射和基因组工程的转化转基因工具。目标的目标侧重于已知或怀疑与之相关的基因的3017蝇基因的资源人类疾病是：（1）生成shrna fly fter库存RNAi，以解决集合中剩余差距，例如尚未涵盖的基因或社区确定的试剂无效；（2）生成sgrna CRISPR的股票（CRISPRKO）和CRISPR激活（CRISPRA）；（3）建立一个试剂工具包结合了Gal4/UAS和第二个二元表达系统（Lexa/Lexaop或QF/Quas），以促进RNAi 和/或CRISPR对两个不同组织中基因表达的控制；（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）表型在我们的“ RSVP Plus”中，有关单个RNAi和SGRNA线质量的新线条和策划信息表型数据库。这些努力的结果将是分配给蝇股的巨大资源布卢明顿果蝇股票中心的社区，该中心将使研究人员轻松击倒，敲出或激活集合所覆盖的基因。当与宽阵列的GAL4线结合时，我们提出的新的Lexa/QF系列集合，该遗传工具包将允许研究人员调节基因在任何给定的阶段和多个时机中的表达。