Increasing the efficiency and targeting range of cytidine base editors through fusion of a single-stranded DNA-binding protein domain.
Nat Cell Biol
; 22(6): 740-750, 2020 06.
Article
em En
| MEDLINE
| ID: mdl-32393889
ABSTRACT
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 ß-haemoglobinopathy, suggesting the therapeutic potential of the improved base editors.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Citidina
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Proteínas de Ligação a DNA
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Rad51 Recombinase
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Sistemas CRISPR-Cas
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Edição de Genes
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Mutação
Limite:
Animals
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Female
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Humans
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article