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

作　　者：Simon Sretenovic Desuo Yin Adam Levav Jeremy D.Selengut Stephen M.Mount Yiping Qi 

机构地区：[1]Department of Plant Science and Landscape Architecture,University of Maryland,College Park,MD 20742,USA [2]Hubei Academy of Agricultural Sciences,Wuhan 430064,China [3]Montgomery Blair High School,Silver Spring,MD 20901,USA [4]Center for Bioinformatics and Computational Biology,University of Maryland,College Park,MD 20742,USA [5]Department of Cell Biology and Molecular Genetics,University of Maryland,College Park,MD 20742,USA [6]Institute for Bioscience and Biotechnology Research,University of Maryland,Rockville,MD 20850,USA

出　　处：《Plant Communications》2021年第2期78-88,共11页植物通讯（英文）

基　　金：supported by startup funds from the University of Maryland,the National Science Foundation Plant Genome Research Program grant(award no.IOS-1758745);the Biotechnology Risk Assessment Grant Program competitive grant(award no.2018-33522-28789)from the U.S.Department of Agriculture.

摘　　要：The most popular CRISPR-SpCas9 systemrecognizes 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 amajor 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 iSpyMac-Cas9 platform.

关 键 词：plant genome editing iSpyMacCas9 PAM cytosine base editing adenine base editing 

分 类 号：Q943.2[生物学—植物学]