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Programmable Single and Multiplex Base-Editing in Using RNA-Guided Cytidine Deaminases.

G3 (Bethesda). 2018-01; 
LiYufeng,MaSanyuan,SunLe,ZhangTong,ChangJiasong,LuWei,ChenXiaoxu,LiuYue,WangXiaogang,ShiRun,ZhaoPing,XiaQin
Products/Services Used Details Operation
Gene Synthesis … DNA encoding rAPOBEC1, the XTEN linker, partial nCas9 with the A840H mutation, 116 Page 7. 7 and UGI was B. mori codon-optimized, synthesized by GenScript service, and then 117 inserted into the pUC57-T-simple plasmid. The complete BE3 vector was constructed 118 … Get A Quote

摘要

Genome editing using standard tools (ZFN, TALEN, and CRISPR/Cas9) rely on double strand breaks to edit the genome. A series of new CRISPR tools that convert cytidine to thymine (C to T) without the requirement for DNA double-strand breaks was developed recently and quickly applied in a variety of organisms. Here, we demonstrate that CRISPR/Cas9-dependent base editor (BE3) converts C to T with a high frequency in the invertebrate silkworm. Using BE3 as a knock-out tool, we inactivated exogenous and endogenous genes through base-editing-induced nonsense mutations with an efficiency of up to 66.2%. Furthermore, genome-scale analysis showed that 96.5% of genes have one or more targetable sites that can ... More

关键词

Bombyx mori,CRISPR/Cas9,DSB-free,base editing,multiplex edi