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Article: Conditional gene inactivation by combining tetracycline-mediated transcriptional repression and auxin-inducible degron-mediated degradation

TitleConditional gene inactivation by combining tetracycline-mediated transcriptional repression and auxin-inducible degron-mediated degradation
Authors
Keywordsmethods
knockout
Inducible degron
Issue Date2019
Citation
Cell Cycle, 2019, v. 18, n. 2, p. 238-248 How to Cite?
AbstractCharacterizing the functions of essential cell cycle control genes requires tight and rapid inducible gene inactivation. Drawbacks of current conditional depletion approaches include slow responses and incomplete depletion. We demonstrated that by integrating the tetracycline-controlled promoter system and the auxin-inducible degron (AID) system together, AID-tagged proteins can be downregulated more efficiently than the individual technology alone. When used in conjunction with CRISPR-Cas9-mediated disruption of the endogenous locus, this system facilitates the analysis of essential genes by allowing rapid and tight conditional depletion, as we have demonstrated using several cell cycle-regulatory genes including cyclin A, CDK2, and TRIP13. The vectors constructed in this study allow expression of AID-fusion proteins under the control of tetracycline-controlled promoters and should be useful in studies requiring rapid and tight suppression of gene expression in mammalian cells.
Persistent Identifierhttp://hdl.handle.net/10722/307260
ISSN
2023 Impact Factor: 3.4
2023 SCImago Journal Rankings: 0.947
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorNg, Lau Yan-
dc.contributor.authorMa, Hoi Tang-
dc.contributor.authorLiu, Julio C.Y.-
dc.contributor.authorHuang, Xiner-
dc.contributor.authorLee, Nelson-
dc.contributor.authorPoon, Randy Y.C.-
dc.date.accessioned2021-11-03T06:22:15Z-
dc.date.available2021-11-03T06:22:15Z-
dc.date.issued2019-
dc.identifier.citationCell Cycle, 2019, v. 18, n. 2, p. 238-248-
dc.identifier.issn1538-4101-
dc.identifier.urihttp://hdl.handle.net/10722/307260-
dc.description.abstractCharacterizing the functions of essential cell cycle control genes requires tight and rapid inducible gene inactivation. Drawbacks of current conditional depletion approaches include slow responses and incomplete depletion. We demonstrated that by integrating the tetracycline-controlled promoter system and the auxin-inducible degron (AID) system together, AID-tagged proteins can be downregulated more efficiently than the individual technology alone. When used in conjunction with CRISPR-Cas9-mediated disruption of the endogenous locus, this system facilitates the analysis of essential genes by allowing rapid and tight conditional depletion, as we have demonstrated using several cell cycle-regulatory genes including cyclin A, CDK2, and TRIP13. The vectors constructed in this study allow expression of AID-fusion proteins under the control of tetracycline-controlled promoters and should be useful in studies requiring rapid and tight suppression of gene expression in mammalian cells.-
dc.languageeng-
dc.relation.ispartofCell Cycle-
dc.subjectmethods-
dc.subjectknockout-
dc.subjectInducible degron-
dc.titleConditional gene inactivation by combining tetracycline-mediated transcriptional repression and auxin-inducible degron-mediated degradation-
dc.typeArticle-
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1080/15384101.2018.1563395-
dc.identifier.pmid30582405-
dc.identifier.pmcidPMC6343694-
dc.identifier.scopuseid_2-s2.0-85059675409-
dc.identifier.volume18-
dc.identifier.issue2-
dc.identifier.spage238-
dc.identifier.epage248-
dc.identifier.eissn1551-4005-
dc.identifier.isiWOS:000456046300008-

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