Eine Plattform für die Wissenschaft: Bauingenieurwesen, Architektur und Urbanistik
Eine Plattform für die Wissenschaft: Bauingenieurwesen, Architektur und Urbanistik
Enhanced Genome Editing Activity with Novel Chimeric ScCas9 Variants in Rice
The Streptococcus canis Cas9 protein (ScCas9) recognizes the NNG protospacer adjacent motif (PAM), offering a wider range of targets than that offered by the commonly used S. pyogenes Cas9 protein (SpCas9). However, both ScCas9 and its evolved Sc++ variant still exhibit low genome editing efficiency in plants, particularly at the less preferred NTG and NCG PAM targets. In this study, a chimeric SpcRN++ variant is engineered by grafting the recognition (REC) domain of SpCas9 into the Sc++ variant, incorporating the R221K/N394K mutations, and retaining the positively charged loop of S. anginosus Cas9. The SpcRN++ variant exhibits a higher genome editing capacity and wider target range than the Sc++ variant in rice protoplasts and stable transgenic plants. Further evidence indicates that nSpcRN++‐based A3A/Y130F and TadA8e exhibit enhanced cytosine and adenine editing efficiency in plants. Finally, herbicide‐resistant rice germplasms are produced by targeting the OsACC gene using nSpcRN++‐based adenine base editors. These results demonstrate that SpcRN++ is a powerful tool for genome editing in plants, and this integrative protein engineering strategy holds promise for engineering other Cas9 proteins.
Enhanced Genome Editing Activity with Novel Chimeric ScCas9 Variants in Rice
The Streptococcus canis Cas9 protein (ScCas9) recognizes the NNG protospacer adjacent motif (PAM), offering a wider range of targets than that offered by the commonly used S. pyogenes Cas9 protein (SpCas9). However, both ScCas9 and its evolved Sc++ variant still exhibit low genome editing efficiency in plants, particularly at the less preferred NTG and NCG PAM targets. In this study, a chimeric SpcRN++ variant is engineered by grafting the recognition (REC) domain of SpCas9 into the Sc++ variant, incorporating the R221K/N394K mutations, and retaining the positively charged loop of S. anginosus Cas9. The SpcRN++ variant exhibits a higher genome editing capacity and wider target range than the Sc++ variant in rice protoplasts and stable transgenic plants. Further evidence indicates that nSpcRN++‐based A3A/Y130F and TadA8e exhibit enhanced cytosine and adenine editing efficiency in plants. Finally, herbicide‐resistant rice germplasms are produced by targeting the OsACC gene using nSpcRN++‐based adenine base editors. These results demonstrate that SpcRN++ is a powerful tool for genome editing in plants, and this integrative protein engineering strategy holds promise for engineering other Cas9 proteins.
Enhanced Genome Editing Activity with Novel Chimeric ScCas9 Variants in Rice
Liang, Zhen (Autor:in) / Wu, Yuqing (Autor:in) / Deng, Shuke (Autor:in) / Wei, Sha (Autor:in) / Zhang, Kai (Autor:in) / Guo, Yingjie (Autor:in)
Advanced Science ; 12
01.02.2025
13 pages
Aufsatz (Zeitschrift)
Elektronische Ressource
Englisch
Enhanced Genome Editing Activity with Novel Chimeric ScCas9 Variants in Rice
| Wiley | 2025
| Wiley | 2024
| Wiley | 2024
THERAPEUTIC USES OF GENOME EDITING WITH CRISPR/Cas SYSTEMS
| Europäisches Patentamt | 2021