Analysis of gene function in somatic mammalian cells using small interfering RNAs.
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.
Elbashir
Sayda M
SM
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, D-37077 Göttingen, Germany.
Harborth
Jens
J
Weber
Klaus
K
Tuschl
Thomas
T
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003599:Cytoskeleton; D004247:DNA; D004305:Dose-Response Relationship, Drug; D004330:Drosophila; D020868:Gene Silencing; D017930:Genes, Reporter; D005821:Genetic Techniques; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D022661:RNA, Untranslated / Q000737:chemistry / Q000235:genetics*; D014162:Transfection
D009841:Oligonucleotides; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D012313:RNA; D004247:DNA; D008156:Luciferases