Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells.
RNA interference (RNAi) was first recognized in Caenorhabditis elegans as a biological response to exogenous double-stranded RNA (dsRNA), which induces sequence-specific gene silencing. RNAi represents a conserved regulatory motif, which is present in a wide range of eukaryotic organisms. Recently, we and others have shown that endogenously encoded triggers of gene silencing act through elements of the RNAi machinery to regulate the expression of protein-coding genes. These small temporal RNAs (stRNAs) are transcribed as short hairpin precursors (approximately 70 nt), processed into active, 21-nt RNAs by Dicer, and recognize target mRNAs via base-pairing interactions. Here, we show that short hairpin RNAs (shRNAs) can be engineered to suppress the expression of desired genes in cultured Drosophila and mammalian cells. shRNAs can be synthesized exogenously or can be transcribed from RNA polymerase III promoters in vivo, thus permitting the construction of continuous cell lines or transgenic animals in which RNAi enforces stable and heritable gene silencing.
Paddison
Patrick J
PJ
Watson School of Biological Sciences, Cold Spring Harbor, New York 11724, USA.
Caudy
Amy A
AA
Bernstein
Emily
E
Hannon
Gregory J
GJ
Conklin
Douglas S
DS
D016475:3T3 Cells; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D017124:Conserved Sequence; D004330:Drosophila; D004722:Endoribonucleases / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D051379:Mice; D012313:RNA / Q000378:metabolism* / Q000494:pharmacology; D012320:RNA Polymerase III / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D043244:Ribonuclease III; D013379:Substrate Specificity; D014162:Transfection
D012333:RNA, Messenger; D012313:RNA; D012320:RNA Polymerase III; D004722:Endoribonucleases; D043244:Ribonuclease III