CORE--HOMOLOGOUS RECOMBINATION FACILITY

核心——同源重组设施

• 批准号: 6594614
• 负责人: ALLAN BRADLEY
• 金额: $ 17.52万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6594614
• 关键词: biomedical facility; congenital heart disorder; embryogenesis; embryonic stem cell; gene targeting; genetic recombination; laboratory mouse; myogenesis

Many of the projects in this program project involve generating mutations in the germ line and somatic tissues of mice by performing homologous recombination, Cre/loxP recombination in ES cells and Cre/loxP recombination in vivo. These procedures for the manipulation of ES cells and the construction of mice are technically demanding and are most efficiently performed in a dedicated laboratory. The various alleles which the core will help to establish in the mouse germ line are described in detail in each of the individual projects. These alleles vary in complexity from "simple" knockouts to more sophisticated alterations generated through successive manipulations of ES cells. For example, in Project V, deletions of about 1Mb will be established on mouse Chromosome 16 using the chromosome engineering techniques pioneered in the laboratory/1. This particular technology requires two gene targeting steps and a Cre-loxP recombination step. Other examples of more sophisticated alleles include flanking the exons of some genes with loxP sites and targeting Cre to specific loci. All of these allele types have been generated previously by the laboratory and therefore this should not present any specific problems. A summary of the types of alleles which will be generated by the core is presented in Figure 1.

该计划项目中的许多项目涉及产生突变 通过同源 ES细胞中的重组，CRE/LOXP重组和CRE/LOXP 体内重组。这些操纵ES细胞的程序 而且，小鼠的建设在技术上是苛刻的，并且是最多的 在专用实验室中有效地进行。各种等位基因 核心将有助于在鼠生系中建立 每个项目中的细节。这些等位基因在 从“简单”淘汰赛到更复杂的更改的复杂性 通过连续对ES细胞的操纵生成。例如，在 项目V，将在鼠标染色体上建立约1MB的删除 16使用染色体工程技术在 实验室/1。这项特定技术需要两个基因靶向步骤 和Cre-loxP重组步骤。其他更复杂的例子 等位基因包括用LOXP位点的某些基因的外显子和 将CRE靶向特定基因座。所有这些等位基因类型已经 实验室先前生成的 提出任何具体问题。摘要的等位基因类型 核心将产生的核心如图1所示。