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A two-plasmid inducible CRISPR/Cas9 genome editing tool for Clostridium acetobutylicum.

J. Microbiol. Methods. 2017; 
Wasels Fran?ois,Jean-Marie Jennifer,Collas Florent,López-Contreras Ana M,Lopes Ferreira Nic
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Nucleic Acid Purification & Analysis A codon-optimised cas9 gene from S. pyogenes flanked by NcoI and SalI was synthesised (Genscript, Supplementary Text) and cloned into a NcoI/SalI-double-digested vector, yielding pCas9con. Get A Quote

摘要

CRISPR/Cas-based genetic engineering has revolutionised molecular biology in both eukaryotes and prokaryotes. Several tools dedicated to the genomic transformation of the Clostridium genus of Gram-positive bacteria have been described in the literature; however, the integration of large DNA fragments still remains relatively limited. In this study, a CRISPR/Cas9 genome editing tool using a two-plasmid strategy was developed for the solventogenic strain Clostridium acetobutylicum ATCC 824. Codon-optimised cas9 from Streptococcus pyogenes was placed under the control of an anhydrotetracycline-inducible promoter on one plasmid, while the gRNA expression cassettes and editing templates were located on a secon... More

关键词

CRISPR/Cas9,Clostridium acetobutylicum,Genome engineering,Metabolic enginee