Laboratory of Combinatorial Genetics and Synthetic Biology, School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
Department of Electrical and Electronic Engineering, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
STAR Protoc. 2021 Jan 9;2(1):100255. doi: 10.1016/j.xpro.2020.100255. eCollection 2021 Mar 19.
The CRISPR-Cas system coupled with Combinatorial Genetics (CombiGEM) enables systematic analysis of high-order genetic perturbations that are important for understanding biological processes and discovering therapeutic target combinations. Here, we present detailed steps and technical considerations for building multiplexed guide RNA libraries and carrying out a combinatorial CRISPR screen in mammalian cells. We also present an analytical pipeline, CombiPIPE, for mapping two- and three-way genetic interactions. For complete details on the use and execution of this protocol, please refer to Zhou et al. (2020).
CRISPR-Cas 系统与组合遗传学（CombiGEM）相结合，使系统分析高阶遗传干扰成为可能，这对于理解生物过程和发现治疗靶组合非常重要。 在这里，我们介绍了构建多路向导 RNA 文库并在哺乳动物细胞中进行组合 CRISPR 筛选的详细步骤和技术注意事项。 我们还提出了一个分析管道 CombiPIPE，用于绘制二项和三项遗传相互作用。 有关此方案的使用和执行的完整详细信息，请参阅 Zhou 等人。 （2020）。
