Heritable and inducible genetic interference by double-stranded RNA encoded by transgenes

Nat Genet. 2000 Feb;24(2):180-3. doi: 10.1038/72850.

Abstract

Double-stranded RNA interference (RNAi) is an effective method for disrupting expression of specific genes in Caenorhabditis elegans and other organisms. Applications of this reverse-genetics tool, however, are somewhat restricted in nematodes because introduced dsRNA is not stably inherited. Another difficulty is that RNAi disruption of late-acting genes has been generally less consistent than that of embryonically expressed genes, perhaps because the concentration of dsRNA becomes lower as cellular division proceeds or as developmental time advances. In particular, some neuronally expressed genes appear refractory to dsRNA-mediated interference. We sought to extend the applicability of RNAi by in vivo expression of heritable inverted-repeat (IR) genes. We assayed the efficacy of in vivo-driven RNAi in three situations for which heritable, inducible RNAi would be advantageous: (i) production of large numbers of animals deficient for gene activities required for viability or reproduction; (ii) generation of large populations of phenocopy mutants for biochemical analysis; and (iii) effective gene inactivation in the nervous system. We report that heritable IR genes confer potent and specific gene inactivation for each of these applications. We suggest that a similar strategy might be used to test for dsRNA interference effects in higher organisms in which it is feasible to construct transgenic animals, but impossible to directly or transiently introduce high concentrations of dsRNA.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans Proteins*
  • Gene Expression Regulation
  • Genes, Helminth*
  • Green Fluorescent Proteins
  • Heat-Shock Proteins / genetics*
  • Helminth Proteins / genetics
  • Luminescent Proteins / genetics
  • Nucleic Acid Conformation
  • Promoter Regions, Genetic
  • RNA, Double-Stranded / chemistry
  • RNA, Double-Stranded / genetics*
  • Recombinant Fusion Proteins / biosynthesis

Substances

  • Caenorhabditis elegans Proteins
  • Heat-Shock Proteins
  • Helminth Proteins
  • Luminescent Proteins
  • RNA, Double-Stranded
  • Recombinant Fusion Proteins
  • hsp-16.2 protein, C elegans
  • Green Fluorescent Proteins