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Strand-preferred base editing of organellar and nuclear genomes using CyDENT

Nat Biotechnol .. 2023-08; 
Jiacheng Hu , Yu Sun , Boshu Li , Zhen Liu , Zhiwei Wang , Qiang Gao , Mengyue Guo , Guanwen Liu , Kevin Tianmeng Zhao , Caixia Gao
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Gene Synthesis Genes encoding the modules for cpCyDENT and part of the mtCyDENT were codon-optimized and synthesized by GenScript. Get A Quote

摘要

Transcription-activator-like effector (TALE)-based tools for base editing of nuclear and organellar DNA rely on double-stranded DNA deaminases, which edit substrate bases on both strands of DNA, reducing editing precision. Here, we present CyDENT base editing, a CRISPR-free, strand-selective, modular base editor. CyDENT comprises a pair of TALEs fused with a FokI nickase, a single-strand-specific cytidine deaminase and an exonuclease to generate a single-stranded DNA substrate for deamination. We demonstrate effective base editing in nuclear, mitochondrial and chloroplast genomes. At certain mitochondrial sites, we show editing efficiencies of 14% and strand specificity of 95%. Furthermore, by exchanging the Cy... More

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