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

文献类型： 外文期刊

作者： Sretenovic, Simon 1 ; Yin, Desuo 1 ; Levav, Adam 1 ; Selengut, Jeremy D. 4 ; Mount, Stephen M. 5 ; Qi, Yiping 1 ;

作者机构： 1.Univ Maryland, Dept Plant Sci & Landscape Architecture, College Pk, MD 20742 USA

2.Hubei Acad Agr Sci, Wuhan 430064, Peoples R China

3.Montgomery Blair High Sch, Silver Spring, MD 20901 USA

4.Univ Maryland, Ctr Bioinformat & Computat Biol, College Pk, MD 20742 USA

5.Univ Maryland, Dept Cell Biol & Mol Genet, College Pk, MD 20742 USA

6.Univ Maryland, Inst Biosci & Biotechnol Res, Rockville, MD 20850 USA

关键词： plant genome editing; iSpyMacCas9; PAM; cytosine base editing; adenine base editing

期刊名称：PLANT COMMUNICATIONS （ 影响因子：8.625； 五年影响因子：8.625 ）

ISSN： 2590-3462

年卷期： 2021 年 2 卷 2 期

页码：

收录情况： SCI

摘要： 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.

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