High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 32661245.
- Also identified by DOI 10.1038/s41467-020-17209-1 and PMC identifier 7359328.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Bacterial Proteins
- CRISPR-Associated Proteins
- CRISPR-Cas Systems
- Endodeoxyribonucleases
- Gene Editing
- Gene Expression Regulation, Leukemic
- Leukemia