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

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 cel...

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Autores principales: Gier, Rodrigo A., Budinich, Krista A., Evitt, Niklaus H., Cao, Zhendong, Freilich, Elizabeth S., Chen, Qingzhou, Qi, Jun, Lan, Yemin, Kohli, Rahul M., Shi, Junwei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359328/
https://www.ncbi.nlm.nih.gov/pubmed/32661245
http://dx.doi.org/10.1038/s41467-020-17209-1
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author Gier, Rodrigo A.
Budinich, Krista A.
Evitt, Niklaus H.
Cao, Zhendong
Freilich, Elizabeth S.
Chen, Qingzhou
Qi, Jun
Lan, Yemin
Kohli, Rahul M.
Shi, Junwei
author_facet Gier, Rodrigo A.
Budinich, Krista A.
Evitt, Niklaus H.
Cao, Zhendong
Freilich, Elizabeth S.
Chen, Qingzhou
Qi, Jun
Lan, Yemin
Kohli, Rahul M.
Shi, Junwei
author_sort Gier, Rodrigo A.
collection PubMed
description 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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spelling pubmed-73593282020-07-20 High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening Gier, Rodrigo A. Budinich, Krista A. Evitt, Niklaus H. Cao, Zhendong Freilich, Elizabeth S. Chen, Qingzhou Qi, Jun Lan, Yemin Kohli, Rahul M. Shi, Junwei Nat Commun Article 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. Nature Publishing Group UK 2020-07-13 /pmc/articles/PMC7359328/ /pubmed/32661245 http://dx.doi.org/10.1038/s41467-020-17209-1 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Gier, Rodrigo A.
Budinich, Krista A.
Evitt, Niklaus H.
Cao, Zhendong
Freilich, Elizabeth S.
Chen, Qingzhou
Qi, Jun
Lan, Yemin
Kohli, Rahul M.
Shi, Junwei
High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening
title High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening
title_full High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening
title_fullStr High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening
title_full_unstemmed High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening
title_short High-performance CRISPR-Cas12a genome editing for combinatorial genetic screening
title_sort high-performance crispr-cas12a genome editing for combinatorial genetic screening
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7359328/
https://www.ncbi.nlm.nih.gov/pubmed/32661245
http://dx.doi.org/10.1038/s41467-020-17209-1
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