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Updated: Dec 15, 2025

Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Published on: September 4, 2019
High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening
Rodrigo A Gier1,2,3,4, Krista A Budinich1,2,3, Niklaus H Evitt1,2,3,5
1Department of Cancer Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA.
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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