Efficient and specific gene knockdown by small interfering RNAs produced in bacteria View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2013-04

AUTHORS

Linfeng Huang, Jingmin Jin, Padraig Deighan, Evgeny Kiner, Larry McReynolds, Judy Lieberman

ABSTRACT

Synthetic small interfering RNAs (siRNAs) are an indispensable tool to investigate gene function in eukaryotic cells and may be used for therapeutic purposes to knock down genes implicated in disease. Thus far, most synthetic siRNAs have been produced by chemical synthesis. Here we present a method to produce highly potent siRNAs in Escherichia coli. This method relies on ectopic expression of p19, an siRNA-binding protein found in a plant RNA virus. When expressed in E. coli, p19 stabilizes an ∼21-nt siRNA-like species produced by bacterial RNase III. When mammalian cells are transfected by them, siRNAs that were generated in bacteria expressing p19 and a hairpin RNA encoding 200 or more nucleotides of a target gene reproducibly knock down target 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. More... »

PAGES

350

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt.2537

DOI

http://dx.doi.org/10.1038/nbt.2537

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1013714451

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/23475073


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