RESUMO
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.
Assuntos
Policetídeos , Streptomyces , Animais , Vias Biossintéticas/genética , Sistemas CRISPR-Cas/genética , Edição de Genes , Policetídeos/metabolismo , RNA Guia de Cinetoplastídeos/genética , RNA Guia de Cinetoplastídeos/metabolismo , Streptomyces/genética , Streptomyces/metabolismoRESUMO
A key goal of modular polyketide synthase (PKS) engineering is to alter polyketide stereochemistry. Here we report that exchanging whole PKS modules is a more productive approach than swapping individual ketoreductase (KR) domains for introducing rare 'A2' and 'B2' stereochemistry into model polyketides, and identify four modular 'biobricks' for such synthetic biology efforts.
