Autologous DNA mobilization and multiplication expedite natural products discovery from bacteria

Science. 2024 Dec 13;386(6727):eabq7333. doi: 10.1126/science.abq7333. Epub 2024 Dec 13.

Abstract

The transmission of antibiotic-resistance genes, comprising mobilization and relocation events, orchestrates the dissemination of antimicrobial resistance. Inspired by this evolutionarily successful paradigm, we developed ACTIMOT, a CRISPR-Cas9-based approach to unlock the vast chemical diversity concealed within bacterial genomes. ACTIMOT enables the efficient mobilization and relocation of large DNA fragments from the chromosome to replicative plasmids within the same bacterial cell. ACTIMOT circumvents the limitations of traditional molecular cloning methods involving handling and replicating large pieces of genomic DNA. Using ACTIMOT, we mobilized and activated four cryptic biosynthetic gene clusters from Streptomyces, leading to the discovery of 39 compounds across four distinct classes. This work highlights the potential of ACTIMOT for accelerating the exploration of biosynthetic pathways and the discovery of natural products.

MeSH terms

  • Biological Products* / metabolism
  • Biosynthetic Pathways
  • CRISPR-Cas Systems*
  • DNA Replication
  • DNA, Bacterial / genetics
  • Drug Discovery
  • Drug Resistance, Bacterial / genetics
  • Genome, Bacterial
  • Multigene Family*
  • Plasmids* / genetics
  • Streptomyces* / genetics
  • Streptomyces* / metabolism

Substances

  • Biological Products
  • DNA, Bacterial