High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening-Reference-Cited by-同舟云学术

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

Published:2020-07-13 Issue:1 Volume:11 Page:
ISSN:2041-1723
Container-title:Nature Communications
language:en
Short-container-title:Nat Commun

Author:

Gier Rodrigo A.^ORCID,Budinich Krista A.,Evitt Niklaus H.^ORCID,Cao Zhendong,Freilich Elizabeth S.,Chen Qingzhou,Qi Jun,Lan Yemin,Kohli Rahul M.^ORCID,Shi Junwei^ORCID

Abstract

AbstractCRISPR-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.

Publisher

Springer Science and Business Media LLC

Subject

General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry

Link

http://www.nature.com/articles/s41467-020-17209-1.pdf

Reference30 articles.

1. Wong, A. S. et al. Multiplexed barcoded CRISPR-Cas9 screening enabled by CombiGEM. Proc. Natl Acad. Sci. USA 113, 2544–2549 (2016).

2. Shen, J. P. et al. Combinatorial CRISPR-Cas9 screens for de novo mapping of genetic interactions. Nat. Methods 14, 573–576 (2017).

3. Han, K. et al. Synergistic drug combinations for cancer identified in a CRISPR screen for pairwise genetic interactions. Nat. Biotechnol. 35, 463–474 (2017).

4. Du, D. et al. Genetic interaction mapping in mammalian cells using CRISPR interference. Nat. Methods 14, 577–580 (2017).

5. Najm, F. J. et al. Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens. Nat. Biotechnol. 36, 179–189 (2018).

Cited by 80 articles. 订阅此论文施引文献订阅此论文施引文献，注册后可以免费订阅5篇论文的施引文献，订阅后可以查看论文全部施引文献

1. CRISPR-Cas assisted diagnostics of plant viruses and challenges;Virology;2024-09

2. The type V effectors for CRISPR/Cas-mediated genome engineering in plants;Biotechnology Advances;2024-09

3. Nucleolytic processing of abasic sites underlies PARP inhibitor hypersensitivity in ALC1-deficient BRCA mutant cancer cells;Nature Communications;2024-07-27

4. AP-1 Mediates Cellular Adaptation and Memory Formation During Therapy Resistance;2024-07-25

5. CRISPR/Cas12a Collateral Cleavage-Driven Transcription Amplification for Direct Nucleic Acid Detection;Analytical Chemistry;2024-07-17