Autologous DNA mobilization and multiplication expedite natural products discovery from bacteria

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.