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Article: Gene down-regulation with short hairpin RNAs and validation of specificity by inducible rescue in mammalian cells

TitleGene down-regulation with short hairpin RNAs and validation of specificity by inducible rescue in mammalian cells
Authors
KeywordsRNA interference
Inducible expression
Short hairpin RNA
Issue Date2010
Citation
Current Protocols in Cell Biology, 2010, suppl. 49, unit 27.2 How to Cite?
AbstractThe principal problem with RNA interference (RNAi) experiments is off-target effects. The most vigorous demonstration of the specificity is the rescue of the RNAi effects with an RNAi-resistant target gene. By combining the expression of short hairpin RNA (shRNA) and rescue cDNA in the same vector, both the validation of shRNA specificity and the generation of shRNA-expressing cell lines can easily be accomplished. If the compensatory cDNA is under the control of an inducible promoter, stable shRNA-expressing cells can be generated before the knockdown phenotypes are studied, by conditionally turning off the rescue protein. The use of model systems is detailed in these protocols. © 2010 by John Wiley & Sons, Inc.
Persistent Identifierhttp://hdl.handle.net/10722/307108
ISSN
2023 SCImago Journal Rankings: 1.872

 

DC FieldValueLanguage
dc.contributor.authorMa, Hoi Tang-
dc.contributor.authorPoon, Randy Y.C.-
dc.date.accessioned2021-11-03T06:21:57Z-
dc.date.available2021-11-03T06:21:57Z-
dc.date.issued2010-
dc.identifier.citationCurrent Protocols in Cell Biology, 2010, suppl. 49, unit 27.2-
dc.identifier.issn1934-2500-
dc.identifier.urihttp://hdl.handle.net/10722/307108-
dc.description.abstractThe principal problem with RNA interference (RNAi) experiments is off-target effects. The most vigorous demonstration of the specificity is the rescue of the RNAi effects with an RNAi-resistant target gene. By combining the expression of short hairpin RNA (shRNA) and rescue cDNA in the same vector, both the validation of shRNA specificity and the generation of shRNA-expressing cell lines can easily be accomplished. If the compensatory cDNA is under the control of an inducible promoter, stable shRNA-expressing cells can be generated before the knockdown phenotypes are studied, by conditionally turning off the rescue protein. The use of model systems is detailed in these protocols. © 2010 by John Wiley & Sons, Inc.-
dc.languageeng-
dc.relation.ispartofCurrent Protocols in Cell Biology-
dc.subjectRNA interference-
dc.subjectInducible expression-
dc.subjectShort hairpin RNA-
dc.titleGene down-regulation with short hairpin RNAs and validation of specificity by inducible rescue in mammalian cells-
dc.typeArticle-
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1002/0471143030.cb2702s49-
dc.identifier.pmid21154550-
dc.identifier.scopuseid_2-s2.0-79953184979-
dc.identifier.issuesuppl. 49-
dc.identifier.spageunit 27.2-
dc.identifier.epageunit 27.2-
dc.identifier.eissn1934-2616-

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