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High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening.

ABSTRACT: CRISPR-based genetic screening has revolutionized cancer drug target discovery, yet reliable, multiplex gene editing to reveal synergies between gene targets remains a major challenge. Here, we present a simple and robust CRISPR-Cas12a-based approach for combinatorial genetic screening in cancer cells. By engineering the CRISPR-AsCas12a system with key modifications to the Cas protein and its CRISPR RNA (crRNA), we can achieve high efficiency combinatorial genetic screening. We demonstrate the performance of our optimized AsCas12a (opAsCas12a) through double knockout screening against epigenetic regulators. This screen reveals synthetic sick interactions between Brd9&Jmjd6, Kat6a&Jmjd6, and Brpf1&Jmjd6 in leukemia cells.

SUBMITTER: Gier RA

PROVIDER: S-EPMC7359328 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening.

Gier Rodrigo A RA Budinich Krista A KA Evitt Niklaus H NH Cao Zhendong Z Freilich Elizabeth S ES Chen Qingzhou Q Qi Jun J Lan Yemin Y Kohli Rahul M RM Shi Junwei J

Nature communications 20200713 1

CRISPR-based genetic screening has revolutionized cancer drug target discovery, yet reliable, multiplex gene editing to reveal synergies between gene targets remains a major challenge. Here, we present a simple and robust CRISPR-Cas12a-based approach for combinatorial genetic screening in cancer cells. By engineering the CRISPR-AsCas12a system with key modifications to the Cas protein and its CRISPR RNA (crRNA), we can achieve high efficiency combinatorial genetic screening. We demonstrate the p ...[more]

PMID: 32661245

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