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

Rodrigo A. Gier(Cancer Research Institute), Krista A. Budinich(Cancer Research Institute), Niklaus H. Evitt(Cancer Research Institute), Zhendong Cao(Cancer Research Institute), Elizabeth Freilich(Cancer Research Institute), Qingzhou Chen(Cancer Research Institute), Jun Qi(Harvard University), Yemin Lan(University of Pennsylvania), Rahul M. Kohli(University of Pennsylvania), Junwei Shi(University of Pennsylvania)
Nature Communications
July 13, 2020
10.1038/s41467-020-17209-1
Cited by 142Open Access
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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.

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