RT Journal Article
SR Electronic
T1 A genome-scale CRISPR interference guide library enables comprehensive phenotypic profiling in yeast
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.11.988105
DO 10.1101/2020.03.11.988105
A1 Nicholas J. McGlincy
A1 Zuriah A. Meacham
A1 Kendra Swain
A1 Ryan Muller
A1 Rachel Baum
A1 Nicholas T. Ingolia
YR 2020
UL http://biorxiv.org/content/early/2020/03/12/2020.03.11.988105.abstract
AB CRISPR/Cas9-mediated transcriptional interference (CRISPRi) enables programmable gene knock-down, yielding interpretable loss-of-function phenotypes for nearly any gene. Effective, inducible CRISPRi has been demonstrated in budding yeast, but no genome-scale guide libraries have been reported. We present a comprehensive yeast CRISPRi library, based on empirical design rules, containing 10 distinct guides for most genes. Competitive growth after pooled transformation revealed strong fitness defects for most essential genes, verifying that the library provides comprehensive genome coverage. We used the relative growth defects caused by different guides targeting essential genes to further refine yeast CRISPRi design rules. In order to obtain more accurate and robust guide abundance measurements in pooled screens, we link guides with random nucleotide barcodes and carry out linear amplification by in vitro transcription. Taken together, we demonstrate a broadly useful platform for comprehensive, high-precision CRISPRi screening in yeast.