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CRISPR-BEST: a highly efficient DSB-free base editor for filamentous actinomycetes

By Yaojun Tong, Helene L. Robertsen, Kai Blin, Andreas K. Klitgaard, Tilmann Weber, Sang Yup Lee

Posted 20 Mar 2019
bioRxiv DOI: 10.1101/582403 (published DOI: 10.1073/pnas.1913493116)

Filamentous actinomycetes serve as major producers of various natural products including antimicrobial compounds. Although CRISPR-Cas9 systems have been developed for more robust genetic manipulations, concerns of genome instability caused by the DNA double-strand breaks (DSB) and the toxicity of Cas9 remain. To overcome these limitations, here we report development of the DSB-free, single-nucleotide resolution genome editing system CRISPR-BEST (CRISPR-Base Editing SysTem). Specifically targeted by an sgRNA, the cytidine deaminase component of CRISPR-BEST efficiently converts C:G to T:A within a window of approximately seven-nucleotides. The system was validated and successfully used in different Streptomyces species.

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