NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for AIDS Research, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, P. R. China.
McGill University AIDS Centre, Lady Davis Institute, Jewish General Hospital, Montreal H3T 1E2, Canada.
STAR Protoc. 2021 Jul 3;2(3):100649. doi: 10.1016/j.xpro.2021.100649. eCollection 2021 Sep 17.
The cyclic GMP-AMP synthase (cGAS) is the principal DNA sensor, which binds DNA and triggers the type I interferon production. We used ISD45 or inactivated Vaccinia Virus (VACV) to stimulate cGAS and monitored cellular localization by immunofluorescence microscopy, Operetta high-content screening, and cytoplasmic/nuclear fractionation. LocNES server was used to predict cGAS nuclear export signal (NES) sequence and characterized the function by mutagenesis. This protocol provides a prototype of cGAS subcellular distribution or the identification of NES in other proteins. For complete details on the use and execution of this protocol, please refer to Sun et al. Sun et al. (2021).
环鸟苷酸-腺苷酸合酶（cGAS）是主要的 DNA 传感器，它可以与 DNA 结合并触发 I 型干扰素的产生。我们使用 ISD45 或已灭活的牛痘病毒（VACV）来刺激 cGAS，并通过免疫荧光显微镜、Operetta 高通量筛选和细胞质/核分馏来监测细胞定位。LocNES 服务器用于预测 cGAS 核输出信号（NES）序列，并通过突变来表征其功能。该方案提供了 cGAS 亚细胞分布或其他蛋白质中 NES 鉴定的原型。有关该方案使用和执行的完整详细信息，请参阅 Sun 等人的研究。Sun 等人。（2021）。
