A genome‐scale yeast library with inducible expression of individual genes
Abstract 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 whic...
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| Format: | Article |
| Language: | English |
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Springer Nature
2021-06-01
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| Series: | Molecular Systems Biology |
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| Online Access: | https://doi.org/10.15252/msb.202110207 |
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| author | Yuko Arita Griffin Kim Zhijian Li Helena Friesen Gina Turco Rebecca Y Wang Dale Climie Matej Usaj Manuel Hotz Emily H Stoops Anastasia Baryshnikova Charles Boone David Botstein Brenda J Andrews R Scott McIsaac |
| author_facet | Yuko Arita Griffin Kim Zhijian Li Helena Friesen Gina Turco Rebecca Y Wang Dale Climie Matej Usaj Manuel Hotz Emily H Stoops Anastasia Baryshnikova Charles Boone David Botstein Brenda J Andrews R Scott McIsaac |
| author_sort | Yuko Arita |
| collection | DOAJ |
| description | Abstract 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, Z3EB42, that gives lower expression than Z3EV, a feature enabling conditional activation and repression of lowly expressed essential genes that grow without inducer in the YETI library. |
| format | Article |
| id | doaj-art-e823bfdfa6794a8b99835ae6fd0838ee |
| institution | Kabale University |
| issn | 1744-4292 |
| language | English |
| publishDate | 2021-06-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-e823bfdfa6794a8b99835ae6fd0838ee2025-08-20T03:43:31ZengSpringer NatureMolecular Systems Biology1744-42922021-06-0117612310.15252/msb.202110207A genome‐scale yeast library with inducible expression of individual genesYuko Arita0Griffin Kim1Zhijian Li2Helena Friesen3Gina Turco4Rebecca Y Wang5Dale Climie6Matej Usaj7Manuel Hotz8Emily H Stoops9Anastasia Baryshnikova10Charles Boone11David Botstein12Brenda J Andrews13R Scott McIsaac14Terrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoCalico Life Sciences LLCTerrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoTerrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoCalico Life Sciences LLCCalico Life Sciences LLCTerrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoTerrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoCalico Life Sciences LLCCalico Life Sciences LLCCalico Life Sciences LLCTerrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoCalico Life Sciences LLCTerrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoCalico Life Sciences LLCAbstract 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, Z3EB42, that gives lower expression than Z3EV, a feature enabling conditional activation and repression of lowly expressed essential genes that grow without inducer in the YETI library.https://doi.org/10.15252/msb.202110207BAR‐seqCRISPRigene overexpressionyeast genomicsyeast mutant array |
| spellingShingle | Yuko Arita Griffin Kim Zhijian Li Helena Friesen Gina Turco Rebecca Y Wang Dale Climie Matej Usaj Manuel Hotz Emily H Stoops Anastasia Baryshnikova Charles Boone David Botstein Brenda J Andrews R Scott McIsaac A genome‐scale yeast library with inducible expression of individual genes Molecular Systems Biology BAR‐seq CRISPRi gene overexpression yeast genomics yeast mutant array |
| title | A genome‐scale yeast library with inducible expression of individual genes |
| title_full | A genome‐scale yeast library with inducible expression of individual genes |
| title_fullStr | A genome‐scale yeast library with inducible expression of individual genes |
| title_full_unstemmed | A genome‐scale yeast library with inducible expression of individual genes |
| title_short | A genome‐scale yeast library with inducible expression of individual genes |
| title_sort | genome scale yeast library with inducible expression of individual genes |
| topic | BAR‐seq CRISPRi gene overexpression yeast genomics yeast mutant array |
| url | https://doi.org/10.15252/msb.202110207 |
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