- High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening.
High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening.
Nature communications (2020-07-15)
Rodrigo A Gier, Krista A Budinich, Niklaus H Evitt, Zhendong Cao, Elizabeth S Freilich, Qingzhou Chen, Jun Qi, Yemin Lan, Rahul M Kohli, Junwei Shi
PMID32661245
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.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
Anti-α-Tubulin antibody, Mouse monoclonal, clone DM1A, purified from hybridoma cell culture