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

被引:11
作者
Yue, Xue [1 ]
Xia, Tianyu [2 ,3 ]
Wang, Shuai [1 ]
Dong, Huijun [1 ]
Li, Yongquan [2 ,3 ]
机构
[1] Liaocheng Univ, Sch Pharm, 1 Hunan Rd, Liaocheng 252000, Shandong, Peoples R China
[2] Zhejiang Univ, Inst Pharmaceut Biotechnol, Sch Med, Affiliated Hosp 1, 866 Yuhangtang Rd, Hangzhou 310058, Zhejiang, Peoples R China
[3] Zhejiang Univ, Affiliated Hosp 1, Sch Med, Res Ctr Clin Pharm, 866 Yuhangtang Rd, Hangzhou 310058, Zhejiang, Peoples R China
关键词
A40926; CRISPR; Cas9; Genome editing; N; gerenzanensis; RecA; GLYCOPEPTIDE ANTIBIOTIC A40926; STREPTOMYCES; BIOSYNTHESIS;
D O I
10.1007/s10529-020-02893-2
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 Actinomyces able to shorten editing time simultaneously.
引用
收藏
页码:1699 / 1706
页数:8
相关论文
共 14 条
  • [1] Genetic manipulation of secondary metabolite biosynthesis for improved production in Streptomyces and other actinomycetes
    Baltz, Richard H.
    [J]. JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 2016, 43 (2-3) : 343 - 370
  • [2] In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway
    Cai, Yuan
    Cheng, Tianlin
    Yao, Yichuan
    Li, Xiao
    Ma, Yuqian
    Li, Lingyun
    Zhao, Huan
    Bao, Jin
    Zhang, Mei
    Qiu, Zilong
    Xue, Tian
    [J]. SCIENCE ADVANCES, 2019, 5 (04):
  • [3] An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting
    Cao, Jian
    Wu, Lizhen
    Zhang, Shang-Min
    Lu, Min
    Cheung, William K. C.
    Cai, Wesley
    Gale, Molly
    Xu, Qi
    Yan, Qin
    [J]. NUCLEIC ACIDS RESEARCH, 2016, 44 (19)
  • [4] Applications of CRISPR/Cas System to Bacterial Metabolic Engineering
    Cho, Suhyung
    Shin, Jongoh
    Cho, Byung-Kwan
    [J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2018, 19 (04)
  • [5] Consequences of Cas9 cleavage in the chromosome of Escherichia coli
    Cui, Lun
    Bikard, David
    [J]. NUCLEIC ACIDS RESEARCH, 2016, 44 (09) : 4243 - 4251
  • [6] Production of the glycopeptide antibiotic A40926 by Nonomuraea sp ATCC 39727:: influence of medium composition in batch fermentation
    Gunnarsson, N
    Bruheim, P
    Nielsen, J
    [J]. JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 2003, 30 (03) : 150 - 156
  • [7] One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces
    Huang, He
    Zheng, Guosong
    Jiang, Weihong
    Hu, Haifeng
    Lu, Yinhua
    [J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA, 2015, 47 (04) : 231 - 243
  • [8] Regulation of a nickel-cobalt efflux system and nickel homeostasis in a soil actinobacterium Streptomyces coelicolor
    Kim, Hae Mi
    Ahn, Bo-Eun
    Lee, Ju-Hyung
    Roe, Jung-Hye
    [J]. METALLOMICS, 2015, 7 (04) : 702 - 709
  • [9] Protoplast preparation and reversion to the normal filamentous growth in antibiotic-producing uncommon actinomycetes
    Marcone, Giorgia Letizia
    Carrano, Lucia
    Marinelli, Flavia
    Beltrametti, Fabrizio
    [J]. JOURNAL OF ANTIBIOTICS, 2010, 63 (02) : 83 - 88
  • [10] Molecular and functional characterization of the Listeria monocytogenes RecA protein: Insights into the homologous recombination process
    Ojha, Debika
    Patil, K. Neelakanteshwar
    [J]. INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 2020, 119