Open AccessA genome‐scale yeast library with inducible expression of individual genes
Author(s) -
Arita Yuko,
Kim Griffin,
Li Zhijian,
Friesen Helena,
Turco Gina,
Wang Rebecca Y,
Climie Dale,
Usaj Matej,
Hotz Manuel,
Stoops Emily H,
Baryshnikova Anastasia,
Boone Charles,
Botstein David,
Andrews Brenda J,
McIsaac R Scott
Publication year - 2021
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.202110207
Subject(s) - griffin , library science , research centre , biology , history , classics , computer science
The ability to switch a gene from off to on and monitor dynamic changes provides a powerful approach for probing gene function and elucidating causal regulatory relationships. Here, we developed and characterized YETI (Yeast Estradiol strains with Titratable Induction), a collection in which > 5,600 yeast genes are engineered for transcriptional inducibility with single‐gene precision at their native loci and without plasmids. Each strain contains SGA screening markers and a unique barcode, enabling high‐throughput genetics. We characterized YETI using growth phenotyping and BAR‐seq screens, and we used a YETI allele to identify the regulon of Rof1, showing that it acts to repress transcription. We observed that strains with inducible essential genes that have low native expression can often grow without inducer. Analysis of data from eukaryotic and prokaryotic systems shows that native expression is a variable that can bias promoter‐perturbing screens, including CRISPRi. We engineered a second expression system, Z 3 EB42, that gives lower expression than Z 3 EV, a feature enabling conditional activation and repression of lowly expressed essential genes that grow without inducer in the YETI library.