Highly efficient genome editing in N. gerenzanensis using an inducible CRISPR/Cas9–RecA system

• Published in: Biotechnology letters Vol. 42; no. 9; pp. 1699 - 1706
• Main Authors: Yue, Xue, Xia, Tianyu, Wang, Shuai, Dong, Huijun, Li, Yongquan
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.09.2020; Springer Nature B.V
• Subjects: Actinobacteria - genetics; Actinomyces; Actinomyces - genetics; Applied Microbiology; Bacterial Proteins - genetics; Biochemistry; Biomedical and Life Sciences; Biotechnology; CRISPR; CRISPR-Cas systems; CRISPR-Cas Systems - genetics; Editing; Escherichia coli - genetics; Gene Editing - methods; Genome editing; Genomes; Homologous recombination; Homology; Life Sciences; Microbiology; mutants; Mutation - genetics; Nonomuraea; Original Research Paper; Rec A Recombinases - genetics; RecA protein; Recombinase; Genome editing; CRISPR/Cas9; A40926; RecA; N. gerenzanensis
• ISSN: 0141-5492; 1573-6776; 1573-6776
• DOI: 10.1007/s10529-020-02893-2

Objective To develop an inducible CRISPR/Cas9–Recombinase A (RecA) system to manipulate genes in Nonomuraea gerenzanensis effectively. Results Compared with traditional homologous recombination, the inducible CRISPR/Cas9 system achieved 68.8% editing efficiency, whereas, with both the inducible Cas9 and the overexpressed RecA, the efficiency of the combined genome editing system reached 100%. The dbv23 -deleted mutant obtained by the inducible CRISPR/Cas9–RecA system was confirmed to produce more A40926 with an approximate yield of 200 mg L −1 than that of around 150 mg L −1 produced by the wild-type strain. Conclusions This inducible CRISPR/Cas9–RecA system was successfully constructed and can be utilized as an efficient genome editing tool for Actinomyce s able to shorten editing time simultaneously.