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Engineering Modular Polyketide Biosynthesis in Streptomyces Using CRISPR/Cas: A Practical Guide

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Engineering Natural Product Biosynthesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2489))

Abstract

The CRISPR/Cas system, which has been widely applied to organisms ranging from microbes to animals, is currently being adapted for use in Streptomyces bacteria. In this case, it is notably applied to rationally modify the biosynthetic pathways giving rise to the polyketide natural products, which are heavily exploited in the medical and agricultural arenas. Our aim here is to provide the potential user with a practical guide to exploit this approach for manipulating polyketide biosynthesis, by treating key experimental aspects including vector choice, design of the basic engineering components, and trouble-shooting.

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Acknowledgments

We acknowledge financial support from the Agence Nationale de la Recherche (grant number ANR-16-CE92-0006-01, PKS STRUCTURE to K.J.W.), the Université de Lorraine, the Centre National de la Recherche Scientifique (CNRS), and the IMPACT Biomolecules project of the Lorraine Université d’Excellence (Investissements d’avenir - ANR 15-004).

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Authors and Affiliations

  1. Molecular and Structural Enzymology Group, UMR 7365 CNRS-UL IMoPA, Lorraine University, Faculté de médecine, Batiment Biopôle, Vandœuvre-lès-Nancy Cedex, France

    Jean-Malo Massicard, Li Su, Christophe Jacob & Kira J. Weissman

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  1. Jean-Malo Massicard
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  2. Li Su
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  3. Christophe Jacob
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  4. Kira J. Weissman
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Correspondence to Christophe Jacob or Kira J. Weissman .

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  1. Department of Chemistry & BioDiscovery Institute, University of North Texas, Denton, USA

    Elizabeth Skellam

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Massicard, JM., Su, L., Jacob, C., Weissman, K.J. (2022). Engineering Modular Polyketide Biosynthesis in Streptomyces Using CRISPR/Cas: A Practical Guide. In: Skellam, E. (eds) Engineering Natural Product Biosynthesis. Methods in Molecular Biology, vol 2489. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2273-5_10

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  • DOI: https://doi.org/10.1007/978-1-0716-2273-5_10

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