Brief Communication | Published:

CRISPR–Cas9 strategy for activation of silent Streptomyces biosynthetic gene clusters

Nature Chemical Biology volume 13, pages 607609 (2017) | Download Citation

Abstract

Here we report an efficient CRISPR–Cas9 knock-in strategy to activate silent biosynthetic gene clusters (BGCs) in streptomycetes. We applied this one-step strategy to activate multiple BGCs of different classes in five Streptomyces species and triggered the production of unique metabolites, including a novel pentangular type II polyketide in Streptomyces viridochromogenes. This potentially scalable strategy complements existing activation approaches and facilitates discovery efforts to uncover new compounds with interesting bioactivities.

  • Compound

    photocyclized alteramide A

  • Compound

    macrolactam

  • Compound

    FR-900098

  • Compound

    3,6,7,9,11-pentahydroxy-3-methyl-3,4-dihydrobenzo[a]tetracene-1,8,13(2H)-trione

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Acknowledgements

We gratefully acknowledge financial support from the National Institutes of Health (GM077596) (H.Z.), the A*STAR Visiting Investigator Program (H.Z.), and the National Research Foundation, Singapore (NRF2013-THE001-094) (M.M.Z., F.T.W., Y.H.L., E.L.A. and H.Z.). NMR data collection at the UIUC IGB Core was funded by NIH (S10-RR028833). Conjugative donor strains of E. coli were gifted by Prof. William Metcalf at UIUC. We thank members of the Zhao laboratory in UIUC, co-workers in MEL and MERL in A*STAR for constructive comments, Dr. Xudong Guan and Dr. Lingyang Zhu from UIUC for assisting with NMR data acquisition, and Dr. Ying Swan Ho from BTI, A*STAR for HRMS data acquisition.

Author information

Author notes

    • Mingzi M Zhang
    •  & Fong Tian Wong

    These authors contributed equally to this work.

Affiliations

  1. Metabolic Engineering Research Laboratory, Science and Engineering Institutes, Agency for Science, Technology, and Research (A*STAR), Singapore.

    • Mingzi M Zhang
    • , Wan Lin Yeo
    • , Ee Lui Ang
    •  & Huimin Zhao
  2. Molecular Engineering Lab, Biomedical Science Institutes, A*STAR, Singapore.

    • Fong Tian Wong
    •  & Elena Heng
  3. Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana–Champaign, Urbana, Illinois, USA.

    • Yajie Wang
    • , Shangwen Luo
    • , Ryan E Cobb
    • , Behnam Enghiad
    •  & Huimin Zhao
  4. Organic Chemistry, Institute of Chemical and Engineering Sciences, A*STAR, Singapore.

    • Yee Hwee Lim

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Contributions

M.M.Z., F.T.W., and H.Z. conceived and designed the research. M.M.Z., F.T.W., Y.W., E.H., W.L.Y., R.E.C., and B.E. performed the molecular biology, conjugation and fermentation experiments. S.L. and Y.H.L. performed structure elucidation of compounds. M.M.Z., F.T.W., E.L.A., and H.Z. analyzed the data. M.M.Z., F.T.W., and H.Z. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Huimin Zhao.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Results, Supplementary Tables 1–8, Supplementary Figure 1–17 and Supplementary Note

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DOI

https://doi.org/10.1038/nchembio.2341

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