Plasmids

  • pRS416-dCas9-Mxi1 + TetR + pRPR1(TetO)-NotI-gRNA: Plasmid for inducible and tunable CRISPR interference in yeast. pRS416 Ura marked Cen/Ars plasmid with dCas9-Mxi1 under Tef1 promoter, and tet-incucible RPR1 promoter with NotI cloning site adjacent to gRNA. Available from AddGene.
  • pWEB436: integrative E. coli-Streptomyces shuttle vector
  • pTARa: a BAC-based S. cerevisiae/E. coli/Streptomyces shuttle capture vector
  • pJWC1: an IncP1-α group broad-host-range cosmid vector.

Strains

  • Our CRISPRi collection contains ~9000 strains representing 1600 genes (~1100 genes essential for fermentative growth and ~500 genes required for respiration). Each archived strain constitutely expresses dCas9-Mxi1 and contains an integrated, sequence-verified gRNA under control of ATc-inducible promoter. Ref: Smith JS et al., Mol Sys Biol (in press). Please contact Bob St. Onge for access.