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dc.contributor.authorHuang, Linfengen_US
dc.contributor.authorJin, Jingminen_US
dc.contributor.authorDeighan, Padraigen_US
dc.contributor.authorKiner, Evgenyen_US
dc.contributor.authorMcReynolds, Larryen_US
dc.contributor.authorLieberman, Judyen_US
dc.date.accessioned2014-03-10T20:32:48Z
dc.date.issued2013en_US
dc.identifier.citationHuang, Linfeng, Jingmin Jin, Padraig Deighan, Evgeny Kiner, Larry McReynolds, and Judy Lieberman. 2013. “Efficient and specific gene knockdown by small interfering RNAs produced in bacteria.” Nature biotechnology 31 (4): 350-356. doi:10.1038/nbt.2537. http://dx.doi.org/10.1038/nbt.2537.en
dc.identifier.issn1087-0156en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:11878821
dc.description.abstractSynthetic small interfering RNAs (siRNAs) are an indispensable tool to investigate gene function in eukaryotic cells1,2 and may be used for therapeutic purposes to knockdown genes implicated in disease3. Thus far, most synthetic siRNAs have been produced by chemical synthesis. Here we present a method to produce highly potent siRNAs in E. coli. This method relies on ectopic expression of p19, a siRNA-binding protein found in a plant RNA virus4, 5. When expressed in E. coli, p19 stabilizes ~21 nt siRNA-like species produced by bacterial RNase III. Transfection of mammalian cells with siRNAs, generated in bacteria expressing p19 and a hairpin RNA encoding 200 or more nucleotides of a target gene, at low nanomolar concentrations reproducibly knocks down gene expression by ~90% without immunogenicity or off-target effects. Because bacterially produced siRNAs contain multiple sequences against a target gene, they may be especially useful for suppressing polymorphic cellular or viral genes.en
dc.language.isoen_USen
dc.relation.isversionofdoi:10.1038/nbt.2537en
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3622153/pdf/en
dash.licenseLAAen_US
dc.titleEfficient and specific gene knockdown by small interfering RNAs produced in bacteriaen
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden
dc.relation.journalNature biotechnologyen
dash.depositing.authorHuang, Linfengen_US
dc.date.available2014-03-10T20:32:48Z
dc.identifier.doi10.1038/nbt.2537*
dash.contributor.affiliatedDeighan, Padraig
dash.contributor.affiliatedKiner, Evgeny
dash.contributor.affiliatedHuang, Linfeng
dash.contributor.affiliatedLieberman, Judy


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