Enhanced genome editing with a Streptococcus equinus Cas9
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 orth...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-02-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-07593-z |
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| author | Jingtong Liu Yao Wang Jingjing Wei Shengzhou Wang Miaomiao Li Zheyong Huang Sufang Zhang Huihui Liu Jinhai Huang Yongming Wang |
| author_facet | Jingtong Liu Yao Wang Jingjing Wei Shengzhou Wang Miaomiao Li Zheyong Huang Sufang Zhang Huihui Liu Jinhai Huang Yongming Wang |
| author_sort | Jingtong Liu |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-28b9fbbf7c824547b61e57d1985b6457 |
| institution | Kabale University |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-28b9fbbf7c824547b61e57d1985b64572025-02-09T12:50:41ZengNature PortfolioCommunications Biology2399-36422025-02-018111110.1038/s42003-025-07593-zEnhanced genome editing with a Streptococcus equinus Cas9Jingtong Liu0Yao Wang1Jingjing Wei2Shengzhou Wang3Miaomiao Li4Zheyong Huang5Sufang Zhang6Huihui Liu7Jinhai Huang8Yongming Wang9Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Zhongshan Hospital, School of Life Sciences, Fudan UniversityCenter for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Zhongshan Hospital, School of Life Sciences, Fudan UniversityCenter for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Zhongshan Hospital, School of Life Sciences, Fudan UniversityCenter for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Zhongshan Hospital, School of Life Sciences, Fudan UniversityCenter for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Zhongshan Hospital, School of Life Sciences, Fudan UniversityCenter for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Zhongshan Hospital, School of Life Sciences, Fudan UniversityKey Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of ForestryNational Permanent Scientific Research Base for Warm Temperate Zone Forestry of Jiulong Mountain in Beijing, Experimental Center of Forestry in North China, CAFEye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University, NHC Key Laboratory of Myopia (Fudan University), Key Laboratory of Myopia, Chinese Academy of Medical SciencesCenter for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Zhongshan Hospital, School of Life Sciences, Fudan UniversityAbstract 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.https://doi.org/10.1038/s42003-025-07593-z |
| spellingShingle | Jingtong Liu Yao Wang Jingjing Wei Shengzhou Wang Miaomiao Li Zheyong Huang Sufang Zhang Huihui Liu Jinhai Huang Yongming Wang Enhanced genome editing with a Streptococcus equinus Cas9 Communications Biology |
| title | Enhanced genome editing with a Streptococcus equinus Cas9 |
| title_full | Enhanced genome editing with a Streptococcus equinus Cas9 |
| title_fullStr | Enhanced genome editing with a Streptococcus equinus Cas9 |
| title_full_unstemmed | Enhanced genome editing with a Streptococcus equinus Cas9 |
| title_short | Enhanced genome editing with a Streptococcus equinus Cas9 |
| title_sort | enhanced genome editing with a streptococcus equinus cas9 |
| url | https://doi.org/10.1038/s42003-025-07593-z |
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