Yeast CRISPRi

This site is a resource for our system for genome-wide CRISPRi screening in budding yeast (S. cerevisiae), described in McGlincy et al., BMC Genomics 2021. This system comprises:

• A guide RNA library with ten guides per gene for most genes

• Inducible CRISPRi implemented by

  • Expression of the dCas9-Mxi effector, and the tetR regulatory protein, from an integrated construct (derived from Smith et al., Genome Biol 2016)

  • A library of episomal plasmids for tet-inducible expression of guide RNAs from our library

• Random nucleotide barcodes linked with each guide RNA

  • Multiple barcodes per guide provide independent measurements of guide effects

  • Linear barcode amplification by in vitro transcription, reducing noise relative to exponential PCR