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

Rodrigo A. Gier, Krista A. Budinich, Niklaus H. Evitt, Zhendong Cao, Elizabeth S. Freilich, Qingzhou Chen, Jun Qi, Yemin Lan, Rahul M. Kohli and Junwei Shi ()
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Rodrigo A. Gier: University of Pennsylvania
Krista A. Budinich: University of Pennsylvania
Niklaus H. Evitt: University of Pennsylvania
Zhendong Cao: University of Pennsylvania
Elizabeth S. Freilich: University of Pennsylvania
Qingzhou Chen: University of Pennsylvania
Jun Qi: Harvard Medical School
Yemin Lan: University of Pennsylvania
Rahul M. Kohli: University of Pennsylvania
Junwei Shi: University of Pennsylvania

Nature Communications, 2020, vol. 11, issue 1, 1-9

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-17209-1

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