Increasing the efficiency and targeting range of cytidine base editors through fusion of a single-stranded DNA-binding protein domain
| Autor: | Honghui Han, Bailian Cai, Yuxuan Wu, Mengjia Hong, Zhiyong Mao, Yifan Huang, Lei Yang, Caiyu Chen, Zhang Xiaohui, Dali Li, Zuozhen Yang, Liang Chen, Meizhen Liu, Ying Zhang, Biyun Zhu, Weishi Yu, Huiying Li, Liren Wang, Mingyao Liu, Shuming Yin |
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| Rok vydání: | 2020 |
| Předmět: |
Protein domain
RAD51 Cytidine medicine.disease_cause Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Protein Domains Genome editing medicine Animals Humans 030304 developmental biology Gene Editing Mice Inbred ICR 0303 health sciences Mutation Chemistry Cas9 Cell Differentiation Promoter Cell Biology Embryo Mammalian Cell biology DNA-Binding Proteins Mice Inbred C57BL Protospacer adjacent motif HEK293 Cells 030220 oncology & carcinogenesis Female Rad51 Recombinase CRISPR-Cas Systems |
| Zdroj: | Nature Cell Biology. 22:740-750 |
| ISSN: | 1476-4679 1465-7392 |
| DOI: | 10.1038/s41556-020-0518-8 |
| Popis: | Cytidine base editors are powerful genetic tools that catalyse cytidine to thymidine conversion at specific genomic loci, and further improvement of the editing range and efficiency is critical for their broader applications. Through insertion of a non-sequence-specific single-stranded DNA-binding domain from Rad51 protein between Cas9 nickase and the deaminases, serial hyper cytidine base editors were generated with substantially increased activity and an expanded editing window towards the protospacer adjacent motif in both cell lines and mouse embryos. Additionally, hyeA3A-BE4max selectively catalysed cytidine conversion in TC motifs with a broader editing range and much higher activity (up to 257-fold) compared with eA3A-BE4max. Moreover, hyeA3A-BE4max specifically generated a C-to-T conversion without inducing bystander mutations in the haemoglobin gamma gene promoter to mimic a naturally occurring genetic variant for amelioration of β-haemoglobinopathy, suggesting the therapeutic potential of the improved base editors. |
| Databáze: | OpenAIRE |
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