Zhu Shiyou, Zhou Yuexin, Wei Wensheng
Biodynamic Optical Imaging Center (BIOPIC), Beijing Advanced Innovation Center for Genomics, Peking-Tsinghua National Institute of Biological Sciences, State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, No. 5 Yiheyuan Rd., Beijing, 100871, China.
Biodynamic Optical Imaging Center (BIOPIC), Beijing Advanced Innovation Center for Genomics, Peking-Tsinghua Center for Life Sciences, State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, No. 5 Yiheyuan Rd., Beijing, 100871, China.
Methods Mol Biol. 2017;1656:175-181. doi: 10.1007/978-1-4939-7237-1_11.
It is highly desirable to identify gene's function in a high-throughput fashion, and the CRISPR/Cas9 system has been harnessed to meet such a need. Here, we describe a general method to generate genome-scale lentiviral single-guide RNA (sgRNA) library and conduct a pooled function-based screening in human cells. This protocol would be of interest to researchers to rapidly identify genes in a variety of biological processes.
以高通量方式鉴定基因功能是非常必要的，并且CRISPR/Cas9系统已被用于满足这一需求。在此，我们描述了一种生成基因组规模的慢病毒单导向RNA（sgRNA）文库并在人类细胞中进行基于功能的混合筛选的通用方法。该方案将引起研究人员的兴趣，以便在各种生物学过程中快速鉴定基因。
