Analysis of gene function in somatic mammalian cells using small interfering RNAs

Methods. 2002 Feb;26(2):199-213. doi: 10.1016/S1046-2023(02)00023-3.


RNA interference (RNAi) is a highly conserved gene silencing mechanism that uses double-stranded RNA (dsRNA) as a signal to trigger the degradation of homologous mRNA. The mediators of sequence-specific mRNA degradation are 21- to 23-nt small interfering RNAs (siRNAs) generated by ribonuclease III cleavage from longer dsRNAs. Twenty-one-nucleotide siRNA duplexes trigger specific gene silencing in mammalian somatic cells without activation of the unspecific interferon response. Here we provide a collection of protocols for siRNA-mediated knockdown of mammalian gene expression. Because of the robustness of the siRNA knockdown technology, genomewide analysis of human gene function in cultured cells has now become possible.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • Cell Line
  • Cytoskeleton
  • DNA
  • Dose-Response Relationship, Drug
  • Drosophila
  • Gene Silencing*
  • Genes, Reporter
  • Genetic Techniques*
  • HeLa Cells
  • Humans
  • Luciferases / metabolism
  • Microscopy, Fluorescence
  • Models, Genetic
  • Molecular Sequence Data
  • Oligonucleotides / chemistry
  • RNA / chemistry*
  • RNA / metabolism
  • RNA, Messenger / metabolism
  • RNA, Small Interfering
  • RNA, Untranslated / chemistry
  • RNA, Untranslated / genetics*
  • Transfection


  • Oligonucleotides
  • RNA, Messenger
  • RNA, Small Interfering
  • RNA, Untranslated
  • RNA
  • DNA
  • Luciferases