CRISPR/Cas9 system is a robust genome editing platform in biotechnology and medicine.However,it generally produces small insertions/deletions(indels,typically 1-3 bp)but rarely induces larger deletions in specific tar...CRISPR/Cas9 system is a robust genome editing platform in biotechnology and medicine.However,it generally produces small insertions/deletions(indels,typically 1-3 bp)but rarely induces larger deletions in specific target sites.Here,we report a cytidine deaminase-Cas9 fusion-induced deletion system(C-DEL)and an adenine deaminase-Cas9 fusion-induced deletion system(A-DEL)by combining Cas9 with rat APOBEC1(r A1)and Tad A 8e,respectively.Both C-DEL and A-DEL improve the efficiency of deletions compared with the conventional Cas9 system in human cells.In addition,the C-DEL system generates a considerable fraction of predictable multinucleotide deletions from 5’-deaminated C bases to the Cas9-cleavage site and increases the proportion of larger deletions at the target loci.Taken together,the CDEL and A-DEL systems provide a practical strategy for producing efficient multinucleotide deletions,expanding the CRISPR/Cas9 toolsets for gene modifications in human cells.展开更多
基金supported by the National Key Research and Development Program of China Stem Cell and Translational Research(2019YFA0110700)the Program for Changjiang Scholars and Innovative Research Team in University(No.IRT_16R32)+1 种基金the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16030501,XDA16030503)Key Research&Development Program of Guangzhou Regenerative Medicine and Health Guangdong Laboratory(2018GZR110104004)。
文摘CRISPR/Cas9 system is a robust genome editing platform in biotechnology and medicine.However,it generally produces small insertions/deletions(indels,typically 1-3 bp)but rarely induces larger deletions in specific target sites.Here,we report a cytidine deaminase-Cas9 fusion-induced deletion system(C-DEL)and an adenine deaminase-Cas9 fusion-induced deletion system(A-DEL)by combining Cas9 with rat APOBEC1(r A1)and Tad A 8e,respectively.Both C-DEL and A-DEL improve the efficiency of deletions compared with the conventional Cas9 system in human cells.In addition,the C-DEL system generates a considerable fraction of predictable multinucleotide deletions from 5’-deaminated C bases to the Cas9-cleavage site and increases the proportion of larger deletions at the target loci.Taken together,the CDEL and A-DEL systems provide a practical strategy for producing efficient multinucleotide deletions,expanding the CRISPR/Cas9 toolsets for gene modifications in human cells.
