Production of functional small interfering RNAs by an amino-terminal deletion mutant of human Dicer.

Published

Journal Article

Although RNA interference (RNAi) functions as a potent antiviral innate-immune response in plants and invertebrates, mammalian somatic cells appear incapable of mounting an RNAi response and few, if any, small interfering RNAs (siRNAs) can be detected. To examine why siRNA production is inefficient, we have generated double-knockout human cells lacking both Dicer and protein kinase RNA-activated. Using these cells, which tolerate double-stranded RNA expression, we show that a mutant form of human Dicer lacking the amino-terminal helicase domain can process double-stranded RNAs to produce high levels of siRNAs that are readily detectable by Northern blot, are loaded into RNA-induced silencing complexes, and can effectively and specifically inhibit the expression of cognate mRNAs. Remarkably, overexpression of this mutant Dicer, but not wild-type Dicer, also resulted in a partial inhibition of Influenza A virus-but not poliovirus-replication in human cells.

Full Text

Duke Authors

Cited Authors

  • Kennedy, EM; Whisnant, AW; Kornepati, AVR; Marshall, JB; Bogerd, HP; Cullen, BR

Published Date

  • December 15, 2015

Published In

Volume / Issue

  • 112 / 50

Start / End Page

  • E6945 - E6954

PubMed ID

  • 26621737

Pubmed Central ID

  • 26621737

Electronic International Standard Serial Number (EISSN)

  • 1091-6490

Digital Object Identifier (DOI)

  • 10.1073/pnas.1513421112

Language

  • eng

Conference Location

  • United States