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Sretenovic, Simon; Yin, Desuo; Levav, Adam; Selengut, Jeremy D.; Mount, Stephen M.; Qi, Yiping
Plant communications, 03/2021, Letnik: 2, Številka: 2Journal Article
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. This study reports the development of an iSpyMacCas9 genome-editing toolbox for targeted mutagenesis, C to T base editing, and A to G base editing at A-rich PAM sites. While the tools are demonstrated in rice, they are expected to aid genome-editing applications in other plant species as well with a broadened targeting scope.
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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in: SICRIS
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