Genome-Wide CRISPR/Cas9 Screening for High-Throughput Functional Genomics in Human Cells

Shiyou Zhu; Yuexin Zhou; Wensheng Wei

doi:10.1007/978-1-4939-7237-1_11

Genome-Wide CRISPR/Cas9 Screening for High-Throughput Functional Genomics in Human Cells

Methods Mol Biol. 2017:1656:175-181. doi: 10.1007/978-1-4939-7237-1_11.

Authors

Shiyou Zhu¹, Yuexin Zhou², Wensheng Wei³

Affiliations

¹ 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.
² 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.
³ 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. [email protected].

PMID: 28808970
DOI: 10.1007/978-1-4939-7237-1_11

Abstract

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.

Keywords: CRISPR-Cas9 system; High-throughput; Knockout; Screening; sgRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems*
Cell Line
Gene Library*
Genome-Wide Association Study / methods*
Genomics / methods*
Humans