Generation of Cre/lox rats

Cre/lox 大鼠的产生

基本信息

批准号：
9359713
负责人：
Qilong Ying
金额：
$ 78.38万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-15 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9359713
关键词：
Biological Biomedical Research CRISPR/Cas technology Cardiac Cardiology Cells Code Communities DNA cassette Deposition Development Disease Disease model Embryo Experimental Models Functional disorder Funding Gene Targeting Generations Genes Genetic Genetic Recombination Goals Human In Situ Individual Institution Knock-in Knock-out LacZ Genes Letters LoxP-flanked allele Lung Mediating Metabolic Metabolism Missouri Modeling Monoclonal Antibody R24 Mus Mutant Strains Mice Neurologic Neurosciences Organ Partner in relationship Pathogenesis Pathologic Processes Pathway interactions Physiological Processes Physiology Point Mutation Process PubMed Pulmonology Rattus Reagent Regulatory Pathway Reporter Research Research Personnel Resource Development Resources Role Site Study models Technology Time Tissues Training United States National Institutes of Health Work base cell type cost cost effective embryonic stem cell homologous recombination human disease in vivo in vivo Model interest knockout gene new technology offspring physiologic model recombinase-mediated cassette exchange tool

项目摘要

ABSTRACT This R24 resource proposal seeks to promote broad and cost-effective usage of rat models through generation and distribution of a panel of popular Cre/lox knockin rats. Rats have long been used as models that better replicate human physiology and pathophysiology than do mice and are the preferred model for the study of many human diseases. Use of rats to model human disease, however, has been limited until recently by the inability to generate germline-competent rat embryonic stem (ES) cells, precluding the use of ES-cell based approaches to produce conditional/inducible knockout rats. In 2010, we generated the first gene knockout rats by homologous recombination-based gene targeting in ES cells. More recently, we have generated a panel of genetically modified rat lines including gene knockout, knockin of point mutation, knockin of reporters, Cre drivers, and floxed rats. A major strength of the ES cell approach is the ability, in conjunction with Cre/lox technology, to generate rats in which genes are inactivated at specific times and/or in specific tissues or organs. We have assembled a strong group of investigators with complementary expertise to develop a panel of universal Cre/lox reporter rats (Aim 1), cell-specific and inducible Cre rats (Aim 2), and floxed rats in which specific genes of interest are flanked by loxP sites (Aim 3). These Cre/lox rats will allow investigators to visualize specific cell types in situ and can also serve as the basis for the generation of conditional/inducible knockout rats. Cre/lox rats generated in this project will be distributed through the Missouri Rat Resource and Research Center (RRRC) (Aim 4). Availability of these rats would combine the significant biological advantages of the rat with the genetic tractability of the mouse, providing investigators with a highly relevant in vivo model with which to study the contributions of specific genes and pathways to pathogenesis of a number of important human diseases.

抽象的该 R24 资源提案旨在通过生成促进大鼠模型的广泛且具有成本效益的使用以及一组流行的 Cre/lox 基因敲入大鼠的分布。长期以来，老鼠一直被用作更好的模型比小鼠复制人类生理学和病理生理学，是研究的首选模型许多人类疾病。然而，直到最近，使用大鼠来模拟人类疾病仍受到限制。无法产生具有种系能力的大鼠胚胎干 (ES) 细胞，从而排除了基于 ES 细胞的使用产生条件/诱导基因敲除大鼠的方法。 2010年，我们培育出第一只基因敲除大鼠通过 ES 细胞中基于同源重组的基因靶向。最近，我们生成了一个面板转基因大鼠品系，包括基因敲除、点突变敲入、报告基因敲入、Cre 司机和老鼠。 ES 细胞方法的主要优势是能够与 Cre/lox 结合技术，产生基因在特定时间和/或在特定组织中失活的大鼠，或器官。我们组建了一支强大的研究人员小组，他们具有互补的专业知识来开发一个小组通用 Cre/lox 报告大鼠（目标 1）、细胞特异性和诱导型 Cre 大鼠（目标 2）和 floxed 大鼠，其中感兴趣的特定基因两侧是 loxP 位点（目标 3）。这些 Cre/lox 大鼠将使研究人员能够原位可视化特定细胞类型，也可以作为生成条件/诱导细胞的基础淘汰老鼠。本项目中产生的 Cre/lox 大鼠将通过密苏里州大鼠资源进行分发研究中心 (RRRC)（目标 4）。这些大鼠的可用性将结合重要的生物学意义大鼠的优点与小鼠的遗传易驯化性，为研究人员提供了高度相关的体内模型，用于研究特定基因和途径对多种疾病发病机制的贡献人类的重要疾病。