CGBE(C-to-G base editor)systems,pivotal components within the base editing arsenal,enable the precise conversion of cytosines to guanines.However,conventional cytidine deaminases possess non-specific single-stranded D...CGBE(C-to-G base editor)systems,pivotal components within the base editing arsenal,enable the precise conversion of cytosines to guanines.However,conventional cytidine deaminases possess non-specific single-stranded DNA binding properties,leading to off-target effects and safety concerns.The Cas-embedding strategy,which involves embedding functional proteins like deaminases within the Cas9 enzyme’s architecture,emerges as a method to mitigate these off-target effects.Our study pioneers the application of the Cas-embedding strategy to CGBE systems,engineering a suite of novel CGBE editors,CE-CGBE.The CE-CGBE that incorporated eA3A,RBMX and Udgx excelled in editing efficiency,editing purity,and indel formation was named HF-CGBE.HF-CGBE showed no significant difference in off-target effects compared to the negative control group for both DNA and RNA.In summary,the novel HF-CGBE editors we propose expand the base editing toolbox and provide therapeutic approaches for related pathogenic mutations.展开更多
基金support from National Natural Science Foundation of China(82302497).
文摘CGBE(C-to-G base editor)systems,pivotal components within the base editing arsenal,enable the precise conversion of cytosines to guanines.However,conventional cytidine deaminases possess non-specific single-stranded DNA binding properties,leading to off-target effects and safety concerns.The Cas-embedding strategy,which involves embedding functional proteins like deaminases within the Cas9 enzyme’s architecture,emerges as a method to mitigate these off-target effects.Our study pioneers the application of the Cas-embedding strategy to CGBE systems,engineering a suite of novel CGBE editors,CE-CGBE.The CE-CGBE that incorporated eA3A,RBMX and Udgx excelled in editing efficiency,editing purity,and indel formation was named HF-CGBE.HF-CGBE showed no significant difference in off-target effects compared to the negative control group for both DNA and RNA.In summary,the novel HF-CGBE editors we propose expand the base editing toolbox and provide therapeutic approaches for related pathogenic mutations.
