U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targeted gene expression in mammalian cells
- PMID: 11981564
- DOI: 10.1038/nbt0502-497
U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targeted gene expression in mammalian cells
Abstract
The first evidence for gene disruption by double-stranded RNA (dsRNA) came from careful analysis in Caenorhabditis elegans. This phenomenon, called RNA interference (RNAi), was observed subsequently in various organisms, including plants, nematodes, Drosophila, and protozoans. Very recently, it has been reported that in mammalian cells, 21- or 22-nucleotide (nt) RNAs with 2-nt 3' overhangs (small inhibitory RNAs, siRNAs) exhibit an RNAi effect. This is because siRNAs are not recognized by the well-characterized host defense system against viral infections, involving dsRNA-dependent inhibition of protein synthesis. However, the current method for introducing synthetic siRNA into cells by lipofection restricts the range of applications of RNAi as a result of the low transfection efficiencies in some cell types and/or short-term persistence of silencing effects. Here, we report a vector-based siRNA expression system that can induce RNAi in mammalian cells. This technical advance for silencing gene expression not only facilitates a wide range of functional analysis of mammalian genes but might also allow therapeutic applications by means of vector-mediated RNAi.
Comment in
-
Expanding small RNA interference.Nat Biotechnol. 2002 May;20(5):446-8. doi: 10.1038/nbt0502-446. Nat Biotechnol. 2002. PMID: 11981553 No abstract available.
Similar articles
-
[RNA interference directed by small hairpin RNA expressed in COS-7 cells].Yi Chuan Xue Bao. 2003 Apr;30(4):295-300. Yi Chuan Xue Bao. 2003. PMID: 12812050 Chinese.
-
Expression of small interfering RNAs targeted against HIV-1 rev transcripts in human cells.Nat Biotechnol. 2002 May;20(5):500-5. doi: 10.1038/nbt0502-500. Nat Biotechnol. 2002. PMID: 11981565
-
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. Methods. 2002. PMID: 12054897
-
RNA interference--significance and applications.Arch Immunol Ther Exp (Warsz). 2005 Jan-Feb;53(1):39-46. Arch Immunol Ther Exp (Warsz). 2005. PMID: 15761375 Review.
-
Genome-wide application of RNAi to the discovery of potential drug targets.FEBS Lett. 2005 Oct 31;579(26):5988-95. doi: 10.1016/j.febslet.2005.08.015. Epub 2005 Aug 22. FEBS Lett. 2005. PMID: 16153642 Review.
Cited by
-
Single siRNA nanocapsules for enhanced RNAi delivery.J Am Chem Soc. 2012 Aug 22;134(33):13542-5. doi: 10.1021/ja304649a. Epub 2012 Aug 7. J Am Chem Soc. 2012. PMID: 22866878 Free PMC article.
-
The expression of ezrin in NPC and its interaction with NGX6, a novel candidate suppressor.Cancer Sci. 2007 Mar;98(3):341-9. doi: 10.1111/j.1349-7006.2007.00410.x. Cancer Sci. 2007. PMID: 17270023 Free PMC article.
-
A novel siRNA validation system for functional screening and identification of effective RNAi probes in mammalian cells.Biochem Biophys Res Commun. 2006 Aug 4;346(3):707-20. doi: 10.1016/j.bbrc.2006.05.164. Epub 2006 Jun 5. Biochem Biophys Res Commun. 2006. PMID: 16793020 Free PMC article.
-
Comparison of different antisense strategies in mammalian cells using locked nucleic acids, 2'-O-methyl RNA, phosphorothioates and small interfering RNA.Nucleic Acids Res. 2003 Jun 15;31(12):3185-93. doi: 10.1093/nar/gkg409. Nucleic Acids Res. 2003. PMID: 12799446 Free PMC article.
-
Inducible shRNA expression for application in a prostate cancer mouse model.Nucleic Acids Res. 2003 Nov 1;31(21):e127. doi: 10.1093/nar/gng127. Nucleic Acids Res. 2003. PMID: 14576327 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials
