Expanding plant genome-editing scope by an engineered iSpyMacCas9 system that targets A-rich PAM sequences.


Journal

Plant communications
ISSN: 2590-3462
Titre abrégé: Plant Commun
Pays: China
ID NLM: 101769147

Informations de publication

Date de publication:
08 03 2021
Historique:
received: 05 06 2020
revised: 12 07 2020
accepted: 20 07 2020
entrez: 26 4 2021
pubmed: 27 4 2021
medline: 27 4 2021
Statut: epublish

Résumé

The most popular CRISPR-SpCas9 system recognizes canonical NGG protospacer adjacent motifs (PAMs). Previously engineered SpCas9 variants, such as Cas9-NG, favor G-rich PAMs in genome editing. In this manuscript, we describe a new plant genome-editing system based on a hybrid iSpyMacCas9 platform that allows for targeted mutagenesis, C to T base editing, and A to G base editing at A-rich PAMs. This study fills a major technology gap in the CRISPR-Cas9 system for editing NAAR PAMs in plants, which greatly expands the targeting scope of CRISPR-Cas9. Finally, our vector systems are fully compatible with Gateway cloning and will work with all existing single-guide RNA expression systems, facilitating easy adoption of the systems by others. We anticipate that more tools, such as prime editing, homology-directed repair, CRISPR interference, and CRISPR activation, will be further developed based on our promising iSpyMacCas9 platform.

Identifiants

pubmed: 33898973
doi: 10.1016/j.xplc.2020.100101
pii: S2590-3462(20)30129-2
pmc: PMC8060698
doi:

Types de publication

Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Pagination

100101

Informations de copyright

© 2020 The Authors.

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Auteurs

Simon Sretenovic (S)

Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA.

Desuo Yin (D)

Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA.
Hubei Academy of Agricultural Sciences, Wuhan 430064, China.

Adam Levav (A)

Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA.
Montgomery Blair High School, Silver Spring, MD 20901, USA.

Jeremy D Selengut (JD)

Center for Bioinformatics and Computational Biology, University of Maryland, College Park, MD 20742, USA.

Stephen M Mount (SM)

Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA.

Yiping Qi (Y)

Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA.
Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD 20850, USA.

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Classifications MeSH