• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>PLoS Genetics >An Inducible and Reversible Mouse Genetic Rescue System
【24h】

An Inducible and Reversible Mouse Genetic Rescue System

机译：可诱导和可逆的小鼠遗传救援系统

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Inducible and reversible regulation of gene expression is a powerful approach for uncovering gene function. We have established a general method to efficiently produce reversible and inducible gene knockout and rescue in mice. In this system, which we named iKO, the target gene can be turned on and off at will by treating the mice with doxycycline. This method combines two genetically modified mouse lines: a) a KO line with a tetracycline-dependent transactivator replacing the endogenous target gene, and b) a line with a tetracycline-inducible cDNA of the target gene inserted into a tightly regulated (TIGRE) genomic locus, which provides for low basal expression and high inducibility. Such a locus occurs infrequently in the genome and we have developed a method to easily introduce genes into the TIGRE site of mouse embryonic stem (ES) cells by recombinase-mediated insertion. Both KO and TIGRE lines have been engineered for high-throughput, large-scale and cost-effective production of iKO mice. As a proof of concept, we have created iKO mice in the apolipoprotein E (ApoE) gene, which allows for sensitive and quantitative phenotypic analyses. The results demonstrated reversible switching of ApoE transcription, plasma cholesterol levels, and atherosclerosis progression and regression. The iKO system shows stringent regulation and is a versatile genetic system that can easily incorporate other techniques and adapt to a wide range of applications.
机译：基因表达的诱导和可逆调节是揭示基因功能的有力方法。我们已经建立了一种通用方法，可以在小鼠中有效产生可逆和可诱导的基因敲除和拯救。在我们称为iKO的系统中，可以通过用强力霉素对小鼠进行治疗来随意打开和关闭目标基因。该方法结合了两种经过基因修饰的小鼠品系：a）KO品系，用四环素依赖性反式激活子替代内源性靶基因，b）品系，将靶基因的四环素诱导型cDNA插入严格调控（TIGRE）基因组位点，提供低基础表达和高诱导性。这样的基因座很少出现在基因组中，我们已经开发出一种通过重组酶介导的插入轻松将基因导入小鼠胚胎干（ES）细胞TIGRE位点的方法。 KO和TIGRE品系均已设计用于高通量，大规模且经济高效的iKO小鼠生产。作为概念的证明，我们在载脂蛋白E（ApoE）基因中创建了iKO小鼠，该基因可进行敏感和定量的表型分析。结果表明ApoE转录，血浆胆固醇水平以及动脉粥样硬化进展和消退的可逆转换。 iKO系统显示出严格的调节作用，是一种通用的遗传系统，可以轻松地整合其他技术并适应广泛的应用。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号