Efficient and specific gene knockdown by small interfering RNAs produced in bacteria.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Huang, L; Jin, J; Deighan, P; Kiner, E; McReynolds, L; Lieberman, J

Published Date

  • April 2013

Published In

Volume / Issue

  • 31 / 4

Start / End Page

  • 350 - 356

PubMed ID

  • 23475073

Pubmed Central ID

  • PMC3622153

Electronic International Standard Serial Number (EISSN)

  • 1546-1696

International Standard Serial Number (ISSN)

  • 1087-0156

Digital Object Identifier (DOI)

  • 10.1038/nbt.2537

Language

  • eng