RRC ID 77292
Author Zhang X, Chen L, Zhu B, Wang L, Chen C, Hong M, Huang Y, Li H, Han H, Cai B, Yu W, Yin S, Yang L, Yang Z, Liu M, Zhang Y, Mao Z, Wu Y, Liu M, Li D.
Title Increasing the efficiency and targeting range of cytidine base editors through fusion of a single-stranded DNA-binding protein domain.
Journal Nat Cell Biol
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 β-haemoglobinopathy, suggesting the therapeutic potential of the improved base editors.
Volume 22(6)
Pages 740-750
Published 2020-6-1
DOI 10.1038/s41556-020-0518-8
PII 10.1038/s41556-020-0518-8
PMID 32393889
MeSH Animals CRISPR-Cas Systems* Cell Differentiation Cytidine / chemistry Cytidine / genetics* DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism* Embryo, Mammalian / cytology Embryo, Mammalian / metabolism Female Gene Editing* HEK293 Cells Humans Mice Mice, Inbred C57BL Mice, Inbred ICR Mutation* Protein Domains Rad51 Recombinase / genetics Rad51 Recombinase / metabolism*
IF 20.042
Human and Animal Cells HUDEP-2(RCB4557)