Alu-linked hairpins efficiently mediate RNA interference with less toxicity than do H1-expressed short hairpin RNAs
- PMID: 16359634
- DOI: 10.1016/j.ab.2005.11.005
Alu-linked hairpins efficiently mediate RNA interference with less toxicity than do H1-expressed short hairpin RNAs
Abstract
RNA interference has become a powerful tool for specific inhibition of gene expression in mammalian cells. Expression constructs allow for the long-term delivery of short interfering RNAs, usually through the expression of Pol III-transcribed hairpins. In some instances, these expression systems have been shown to have side effects, including induction of the interferon response and cytotoxicity. Here we demonstrate that H1-expressed hairpins, as well as the cloning vector, reduce the plating efficiency of HeLa cells. This toxicity is abrogated by coexpression of the hairpin in the same transcript as a human Alu repetitive element. These Alu-linked hairpins retain the ability to knock down expression of target mRNAs. This modification, which we term SINE (short interspersed repetitive element)-enhanced short hairpin RNA, provides an alternative expression system for hairpins with reduced side effects.
Similar articles
-
Oligonucleotide sequences forming short self-complimentary hairpins can expedite the down-regulation of Coprinopsis cinerea genes.J Microbiol Methods. 2008 Oct;75(2):205-8. doi: 10.1016/j.mimet.2008.06.006. Epub 2008 Jun 17. J Microbiol Methods. 2008. PMID: 18616966
-
Transposon-based RNAi delivery system for generating knockdown cell lines.Biochem Biophys Res Commun. 2004 Apr 9;316(3):643-50. doi: 10.1016/j.bbrc.2004.02.090. Biochem Biophys Res Commun. 2004. PMID: 15033448
-
A tightly regulated and reversibly inducible siRNA expression system for conditional RNAi-mediated gene silencing in mammalian cells.J Gene Med. 2007 Jul;9(7):620-34. doi: 10.1002/jgm.1048. J Gene Med. 2007. PMID: 17486668
-
Expressing short hairpin RNAs in vivo.Nat Methods. 2006 Sep;3(9):689-95. doi: 10.1038/nmeth927. Nat Methods. 2006. PMID: 16929313 Review.
-
Useful 'junk': Alu RNAs in the human transcriptome.Cell Mol Life Sci. 2007 Jul;64(14):1793-800. doi: 10.1007/s00018-007-7084-0. Cell Mol Life Sci. 2007. PMID: 17514354 Free PMC article. Review.
Cited by
-
Safe, long-term hepatic expression of anti-HCV shRNA in a nonhuman primate model.Mol Ther. 2012 Sep;20(9):1737-49. doi: 10.1038/mt.2012.119. Epub 2012 Jun 26. Mol Ther. 2012. PMID: 22735378 Free PMC article.
-
Transposable element insertions have strongly affected human evolution.Proc Natl Acad Sci U S A. 2010 Nov 16;107(46):19945-8. doi: 10.1073/pnas.1014330107. Epub 2010 Nov 1. Proc Natl Acad Sci U S A. 2010. PMID: 21041622 Free PMC article.
-
Argonaute proteins are key determinants of RNAi efficacy, toxicity, and persistence in the adult mouse liver.J Clin Invest. 2010 Sep;120(9):3106-19. doi: 10.1172/JCI43565. Epub 2010 Aug 9. J Clin Invest. 2010. PMID: 20697157 Free PMC article.
-
The L1 retrotransposition assay: a retrospective and toolkit.Methods. 2009 Nov;49(3):219-26. doi: 10.1016/j.ymeth.2009.04.012. Epub 2009 May 4. Methods. 2009. PMID: 19398011 Free PMC article. Review.
-
L1 mobile element expression causes multiple types of toxicity.Gene. 2008 Aug 1;419(1-2):75-81. doi: 10.1016/j.gene.2008.04.013. Epub 2008 May 3. Gene. 2008. PMID: 18555620 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources