Engineer chimeric Cas9 to expand PAM recognition based on evolutionary information

Dacheng Ma, Zhimeng Xu, Zhaoyu Zhang, Xi Chen, Xiangzhi Zeng, Yiyang Zhang, Tingyue Deng, Mengfei Ren, Zheng Sun, Rui Jiang and Zhen Xie ()
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Dacheng Ma: Tsinghua University
Zhimeng Xu: Tsinghua University
Zhaoyu Zhang: Tsinghua University
Xi Chen: Tsinghua University
Xiangzhi Zeng: Tsinghua University
Yiyang Zhang: Tsinghua University
Tingyue Deng: Tsinghua University
Mengfei Ren: Tsinghua University
Zheng Sun: Tsinghua University
Rui Jiang: Tsinghua University
Zhen Xie: Tsinghua University

Nature Communications, 2019, vol. 10, issue 1, 1-9

Abstract: Abstract Although Cas9 nucleases are remarkably diverse in microorganisms, the range of genomic sequences targetable by a CRISPR/Cas9 system is restricted by the requirement of a short protospacer adjacent motif (PAM) at the target site. Here, we generate a group of chimeric Cas9 (cCas9) variants by replacing the key region in the PAM interaction (PI) domain of Staphylococcus aureus Cas9 (SaCas9) with the corresponding region in a panel of SaCas9 orthologs. By using a functional assay at target sites with different nucleotide recombinations at PAM position 3–6, we identify several cCas9 variants with expanded recognition capability at NNVRRN, NNVACT, NNVATG, NNVATT, NNVGCT, NNVGTG, and NNVGTT PAM sequences. In summary, we provide a panel of cCas9 variants accessible up to 1/4 of all the possible genomic targets in mammalian cells.

Date: 2019
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-08395-8

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DOI: 10.1038/s41467-019-08395-8

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