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资源提案旨在通过一代来促进对老鼠模型的广泛和成本效益的用法以及一组流行的Cre/Lox敲击大鼠的分布。老鼠长期以来一直被用作模型复制人类的生理学和病理生理学比小鼠是研究的首选模型许多人类疾病。然而，直到最近，使用大鼠对人类疾病进行建模。无法产生具有生殖线的大鼠胚胎茎（ES）细胞，从而排除了基于ES细胞的使用产生条件/诱导敲除大鼠的方法。 2010年，我们产生了第一个基因敲除大鼠通过在ES细胞中基于基于重组的基因靶向。最近，我们生成了一个小组转基因的大鼠线，包括基因敲除，点突变的敲除，报告者的敲门蛋白，CRE 驾驶员和flox的大鼠。 ES细胞方法的主要优势是与CRE/LOX结合的能力技术，生成大鼠，其中在特定时间和/或特定组织中灭活基因或器官。我们已经组建了一个具有互补专业知识的强大研究人员，以开发一个小组通用CRE/LOX报告基因大鼠（AIM 1），细胞特异性和诱导CRE大鼠（AIM 2）和Floxed大鼠，其中感兴趣的特定基因是LOXP位点的两侧（AIM 3）。这些CRE/LOX大鼠将允许调查人员可视化特定的细胞类型原位，也可以作为有条件/诱导的生成的基础敲除老鼠。该项目中生成的CRE/LOX大鼠将通过密苏里州的Rations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大鼠具有小鼠遗传障碍性的优势，为研究人员提供了高度相关的研究特定基因的贡献和途径对数量的发病机理的贡献的体内模型重要的人类疾病。