A genome-scale yeast library with inducible expression of individual genes.
BAR-seq
CRISPRi
gene overexpression
yeast genomics
yeast mutant array
Journal
Molecular systems biology
ISSN: 1744-4292
Titre abrégé: Mol Syst Biol
Pays: England
ID NLM: 101235389
Informations de publication
Date de publication:
06 2021
06 2021
Historique:
revised:
27
04
2021
received:
06
01
2021
accepted:
30
04
2021
entrez:
7
6
2021
pubmed:
8
6
2021
medline:
14
1
2022
Statut:
ppublish
Résumé
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
Identifiants
pubmed: 34096681
doi: 10.15252/msb.202110207
pmc: PMC8182650
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e10207Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM046406
Pays : United States
Organisme : CIHR
ID : FDN-143264
Pays : Canada
Organisme : CIHR
ID : FDN-143265
Pays : Canada
Informations de copyright
© 2021 The Authors. Published under the terms of the CC BY 4.0 license.
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