重新利用CRISPR作为一种以RNA为导向的平台,用于对基因表达的特定序列控制
Lei S Qi1, Matthew H Larson, Luke A Gilbert
1UCSF Center for Systems and Synthetic Biology, University of California, San Francisco, San Francisco, CA 94158, USA. stanley.qi@ucsf.edu
Cell
|March 5, 2013
概括
我们开发了CRISPR干扰 (CRISPRi),这是一种新的基因编辑工具,使用非活化的Cas9蛋白来精确控制基因表达. 这个系统有效地抑制细菌中的基因,并显示出在哺乳动物细胞中使用的潜力.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物技术是生物技术.
背景情况:
- 全基因组的基因调节对于理解和设计细胞功能至关重要.
- 克里斯普尔系统为精确的DNA向提供了强大的工具.
研究的目的:
- 利用CRISPR技术开发一种用于向基因调节的新方法.
- 为了证明这种系统对基因抑制的有效性和特异性.
主要方法:
- 利用催化死Cas9 (dCas9) 蛋白质,由RNA指导,创建一个DNA识别复合体.
- 在大肠杆菌中与指导RNA共同表达dCas9,以干扰转录过程.
- 评估基因抑制效率和非目标效应.
主要成果:
- 克里斯普尔干扰 (CRISPRi) 在大肠杆菌中有效抑制向基因表达,没有可检测的非向效应.
- 克里斯皮尔系统证明了其能够同时抑制多个基因的能力.
- 该系统的效应被证明是可逆的,并且可以适应哺乳动物细胞中的基因抑制.
结论:
- 克里斯普尔干扰提供了一个简单而有效的RNA引导的DNA识别平台,用于选择性基因扰乱.
- 这项技术使得对基因表达进行全基因组控制,用于研究和工程应用.
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