CRISPR/Cas9-mediated genome editing

Can be used for precise CRISPR/Cas9-mediated genome engineering of Streptomyces strains. Mutations made have ranged from 1-100kbp deletions, precise codon changes to alter amino acids and insertions to add Flag-tags to proteins encoded at their native loci. Several different plasmids have been constructed and published and are listed below. Click the links to find out more.

pCRISPR-Cas9

pCRISPomyces2

For a comprahensive guide for CRISPR/Cas knockouts in Streptomyces using pCRISPomyces-2 see the link below to the 'Hutchings Lab protocol for generating CRISPR/Cas Knockouts using pCRISPomyces-2', written by Dr. Rebecca Devine. [Hutchings Lab protocol for generating CRISPR/Cas Knockouts using pCRISPomyces-2]