Hemphill James, Borchardt Erin K, Brown Kalyn, Asokan Aravind, Deiters Alexander
†Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States.
J Am Chem Soc. 2015 May 6;137(17):5642-5. doi: 10.1021/ja512664v. Epub 2015 Apr 23.
The CRISPR/Cas9 system has emerged as an important tool in biomedical research for a wide range of applications, with significant potential for genome engineering and gene therapy. In order to achieve conditional control of the CRISPR/Cas9 system, a genetically encoded light-activated Cas9 was engineered through the site-specific installation of a caged lysine amino acid. Several potential lysine residues were identified as viable caging sites that can be modified to optically control Cas9 function, as demonstrated through optical activation and deactivation of both exogenous and endogenous gene function.
CRISPR/Cas9系统已成为生物医学研究中一种重要工具，可用于广泛应用，在基因组工程和基因治疗方面具有巨大潜力。为了实现对CRISPR/Cas9系统的条件性控制，通过定点安装一个笼化赖氨酸氨基酸，构建了一种基因编码的光激活Cas9。已鉴定出几个潜在的赖氨酸残基作为可行的笼化位点，可对其进行修饰以光学控制Cas9功能，这已通过外源和内源基因功能的光激活和失活得到证明。
