Genome Editing of Corynebacterium glutamicum Using CRISPR-Cpf1 System.
Methods in molecular biology (Clifton, N.J.)(2022)
摘要
Corynebacterium glutamicum, as an important microbial chassis, has great potential in industrial application. However, complicated genetic modification is severely slowed by lack of efficient genome editing tools. The Streptococcus pyogenes (Sp) CRISPR-Cas9 system has been verified as a very powerful tool for mediating genome alteration in many microorganisms but cannot work well in C. glutamicum. We recently developed two Francisella novicida (Fn) CRISPR-Cpf1 assisted systems for genome editing via homologous recombination in C. glutamicum. Here, we describe the protocols and demonstrated that N iterative rounds of genome editing can be achieved in 3 N + 4 or 3 N + 2 days, respectively.
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关键词
CRISPR-Cpf1 system,Corynebacterium glutamicum,Donor DNA,Genome editing,Homologous recombination,ssDNA
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