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Gene knockdown by ecdysone-based inducible RNAi in stable mammalian cell lines

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dc.contributor.authorRangasamy, Danny
dc.contributor.authorTremethick, David
dc.contributor.authorGreaves, Ian
dc.date.accessioned2015-12-08T22:23:09Z
dc.date.issued2008
dc.date.updated2015-12-08T08:50:02Z
dc.description.abstractRNA interference (RNAi) is a powerful tool for the functional analysis of essential genes in the mammalian genome. Here, we present a simple ecdysone-based inducible RNAi approach that allows high induction and adjustable control of short hairpin RNA (shRNA) expression for silencing gene expression in a wide range of mammalian cells. This protocol describes the following: the design and cloning of inducible shRNA; testing and validation of gene knockdown; and methodology for establishing stable cell lines. This step-by-step protocol offers a quick and cost-effective approach for addressing the function of genes essential for cell cycle regulation and development and can be completed in less than 6 weeks.
dc.identifier.issn1750-2799
dc.identifier.urihttp://hdl.handle.net/1885/32738
dc.publisherNature Publishing Group
dc.sourceNature Protocols
dc.subjectKeywords: ecdysone; ponasterone A; retinoid X receptor; short hairpin RNA; animal cell; article; cell cycle regulation; cell maturation; expression vector; gene expression; gene function; gene induction; gene sequence; gene silencing; human; human cell; mammal cell
dc.titleGene knockdown by ecdysone-based inducible RNAi in stable mammalian cell lines
dc.typeJournal article
local.bibliographicCitation.issue1
local.bibliographicCitation.lastpage88
local.bibliographicCitation.startpage79
local.contributor.affiliationRangasamy, Danny, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationTremethick, David, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationGreaves, Ian, College of Medicine, Biology and Environment, ANU
local.contributor.authoruidRangasamy, Danny, u4020234
local.contributor.authoruidTremethick, David, u9100316
local.contributor.authoruidGreaves, Ian, u4024978
local.description.embargo2037-12-31
local.description.notesImported from ARIES
local.identifier.absfor060199 - Biochemistry and Cell Biology not elsewhere classified
local.identifier.ariespublicationu4020362xPUB95
local.identifier.citationvolume3
local.identifier.doi10.1038/nprot.2007.456
local.identifier.scopusID2-s2.0-38349135364
local.identifier.thomsonID000253140400009
local.type.statusPublished Version

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