Quantitative CRISPR interference screens in yeast identify chemical-genetic interactions and new rules for guide RNA design.

ABSTRACT:

Background

Genome-scale CRISPR interference (CRISPRi) has been used in human cell lines; however, the features of effective guide RNAs (gRNAs) in different organisms have not been well characterized. Here, we define rules that determine gRNA effectiveness for transcriptional repression in Saccharomyces cerevisiae.

Results

We create an inducible single plasmid CRISPRi system for gene repression in yeast, and use it to analyze fitness effects of gRNAs under 18 small molecule treatments. Our approach correctly identifies previously described chemical-genetic interactions, as well as a new mechanism of suppressing fluconazole toxicity by repression of the ERG25 gene. Assessment of multiple target loci across treatments using gRNA libraries allows us to determine generalizable fe

SUBMITTER: Smith JD 

PROVIDER: S-EPMC4784398 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature