Generation of dual-gRNA library for combinatorial CRISPR screening of synthetic lethal gene pairs

Shan Tang, Xue Wu, Jinghui Liu, Qiongsi Zhang, Xinyi Wang, Shuai Shao, Birkan Gokbag, Kunjie Fan, Xiaoqi Liu, Fuhai Li, Lijun Cheng, Lang Li

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Combinatorial CRISPR screening is useful for investigating synthetic lethality (SL) gene pairs. Here, we detail the steps for dual-gRNA library construction, with the introduction of two backbones, LentiGuide_DKO and LentiCRISPR_DKO. We describe steps for in vitro screening with 22Rv1-Cas9 and SaOS2-Cas9 cells followed by sequencing and data analysis. By introducing two backbones, we optimized the library construction process, facilitated standard pair-end sequencing, and provided options of screening on cells with or without modification of Cas9 expression.

Original languageEnglish
Article number101556
JournalSTAR Protocols
Volume3
Issue number3
DOIs
StatePublished - Sep 16 2022

Keywords

  • Biotechnology and bioengineering
  • CRISPR
  • High Throughput Screening
  • Molecular Biology

Fingerprint

Dive into the research topics of 'Generation of dual-gRNA library for combinatorial CRISPR screening of synthetic lethal gene pairs'. Together they form a unique fingerprint.

Cite this