Enhanced genome editing with a Streptococcus equinus Cas9

A large number of SpCas9 orthologs has been computationally identified, but their genome editing potential remains largely unknown. In this study, a GFP-activation assay was used to screen a panel of 18 SpCas9 orthologs, ten of which demonstrated activity in human cells. Notably, these orthologs had a preference for purine-rich PAM sequences. Four of the tested orthologs displayed enhanced specificity compared to SpCas9. Of particular interest is SeqCas9, which recognizes a simple NNG PAM and displays activity and specificity comparable to SpCas9-HF1. In addition, SeqCas9 exhibits superior base editing efficiency compared to SpCas9-NG and SpCas9-NRRH at multiple endogenous loci. This research sheds light on the diversity of SpCas9 orthologs and their potential for specific and efficient genome editing, especially in cases involving base editing.

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Publication Details

Journal
Communications Biology
Published
2025-02-07
DOI
https://doi.org/10.1038/s42003-025-07593-z
Citations
2
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
1.61

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article

Enhanced genome editing with a Streptococcus equinus Cas9

Zheyong Huang, Jingtong Liu, Jinhai Huang, Shengzhou Wang et al.
2 citations
Communications Biology
CRISPR and Genetic Engineering
1.61
article

Enhanced genome editing with a Streptococcus equinus Cas9

Zheyong Huang, Jingtong Liu, Jinhai Huang, Shengzhou Wang, Yao Wang, Miaomiao Li, Yongming Wang, Huihui Liu, Sufang Zhang, Jingjing Wei
article en
2 citations

Abstract

A large number of SpCas9 orthologs has been computationally identified, but their genome editing potential remains largely unknown. In this study, a GFP-activation assay was used to screen a panel of 18 SpCas9 orthologs, ten of which demonstrated activity in human cells. Notably, these orthologs had a preference for purine-rich PAM sequences. Four of the tested orthologs displayed enhanced specificity compared to SpCas9. Of particular interest is SeqCas9, which recognizes a simple NNG PAM and displays activity and specificity comparable to SpCas9-HF1. In addition, SeqCas9 exhibits superior base editing efficiency compared to SpCas9-NG and SpCas9-NRRH at multiple endogenous loci. This research sheds light on the diversity of SpCas9 orthologs and their potential for specific and efficient genome editing, especially in cases involving base editing.

Communications BiologyVol. 8(1)
Chinese Academy of Medical Sciences & Peking Union Medical College (CN), Fudan University (CN), Shanghai Micro Satellite Engineering Center (CN), Huadong Hospital (CN), Eye & ENT Hospital of Fudan University (CN), Zhongshan Hospital (CN), Pudong Medical Center (CN), Chinese Academy of Forestry (CN), Shanghai Fudan Microelectronics (China) (CN), Institute of Forest Ecology, Environment and Protection (CN), Experimental Center of Forestry in North China (CN), Shanghai Clinical Research Center (CN)
National Natural Science Foundation of China, Fudan University, National Key Research and Development Program of China
Zero hunger
Openalex Percentile: Top 19%
CRISPR and Genetic Engineering
1.61
DOIOpen Access PDFOpenAlex
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