CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, P. R. China.
Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, P.R. China.
STAR Protoc. 2021 Jan 27;2(1):100307. doi: 10.1016/j.xpro.2021.100307. eCollection 2021 Mar 19.
We present a detailed protocol for gene editing in adipocytes using the CRISPR-Cas technology. This protocol describes sgRNA design, preparation of lentiCRISPR-sgRNA vectors, functional validation of sgRNAs, preparation of lentiviruses, and lentiviruses transduction in adipocytes. Moreover, an optimized method of gene editing using the lentiCRISPRv2 vector expressing two sgRNAs targeting two different genes has also been described. For complete details on the use and execution of this protocol, please refer to Qiu et al. (2020).
我们介绍了一种使用 CRISPR-Cas 技术在脂肪细胞中进行基因编辑的详细方案。本方案描述了 sgRNA 的设计、lentiCRISPR-sgRNA 载体的制备、sgRNA 的功能验证、慢病毒的制备以及慢病毒在脂肪细胞中的转导。此外，还描述了一种使用靶向两个不同基因的两个 sgRNA 的 lentiCRISPRv2 载体进行基因编辑的优化方法。有关此方案使用和执行的完整详细信息，请参阅 Qiu 等人。（2020 年）。
