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

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. © 2022 The Authors.

Citation

Shan Tang, Xue Wu, Jinghui Liu, Qiongsi Zhang, Xinyi Wang, Shuai Shao, Birkan Gokbag, Kunjie Fan, Xiaoqi Liu, Fuhai Li, Lijun Cheng, Lang Li. Generation of dual-gRNA library for combinatorial CRISPR screening of synthetic lethal gene pairs. STAR protocols. 2022 Sep 16;3(3):101556

Mesh Tags

Substances

PMID: 36060092

search → result