Generation of Cre/lox rats

Cre/lox 大鼠的产生

• 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
• 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

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.

摘要 这份R24资源提案旨在通过以下方式促进大鼠模型的广泛和经济高效的使用 以及一组受欢迎的Cre/lox敲门鼠的分布。长期以来，老鼠一直被用作更好的模型 比小鼠更能复制人的生理学和病理生理学，是研究糖尿病的首选模型 许多人类疾病。然而，直到最近，使用老鼠来模拟人类疾病一直受到限制 不能产生具有生殖系能力的大鼠胚胎干细胞，排除了基于ES细胞的使用 产生条件性/诱导性基因敲除大鼠的方法。2010年，我们培育出了第一只基因敲除大鼠 通过在ES细胞中基于同源重组的基因打靶。最近，我们生成了一个小组 转基因大鼠品系包括基因敲除、点突变敲除、报告基因敲除、Cre 司机和被鞭打的老鼠。ES细胞方法的一个主要优点是能够与CRE/LOX相结合 技术，以产生在特定时间和/或特定组织中基因失活的大鼠 器官。我们已经组建了一支强大的调查小组，他们拥有互补的专业知识，以开发一个小组 通用Cre/lox报告大鼠(目标1)，细胞特异性和诱导性Cre大鼠(目标2)，以及 特定的感兴趣基因位于loxP位点的两侧(目标3)。这些CRE/LOX大鼠将允许调查人员 在原位观察特定的细胞类型，也可以作为产生条件/可诱导的细胞的基础 淘汰鼠。在这个项目中产生的CRE/LOX老鼠将通过密苏里州老鼠资源和 研究中心(RRRC)(目标4)。这些老鼠的可获得性将结合重要的生物学 大鼠的优势与小鼠的遗传易感性，为研究人员提供了一个高度相关的研究领域 体内模型，用来研究特定基因和途径在许多疾病发病中的作用 重要的人类疾病。