Nature Biotechnology 24, 566 - 571 (2006)
Published online: 30 April 2006; | doi:10.1038/nbt1206
Stable expression of shRNAs in human CD34+ progenitor cells can avoid induction of interferon responses to siRNAs in vitroMarjorie A Robbins1, 5, Mingjie Li1, Irene Leung2, Haitang Li1, Doris V Boyer2, Yong Song2, Mark A Behlke3
& John J Rossi1, 41
Division of Molecular Biology, Beckman Research Institute, City of Hope, Duarte, California 91010, USA. 2
Biomedical Research Division, Beckman Coulter, Inc., Fullerton, California 92834, USA. 3
Integrated DNA Technologies, Coralville, Iowa 52241, USA. 4
Division of Molecular Biology, Graduate School of Biological Sciences, Beckman Research Institute, City of Hope, 1450 E. Duarte Rd., Duarte, California 91010. 5
Present address: Protiva Biotherapeutics, Inc., 100-3480 Gilmore Way, Burnaby, British Columbia V5G 4Y1, Canada.
Correspondence should be addressed to John J Rossi jrossi@coh.org RNA interference occurs when cytoplasmic small interfering RNAs (siRNAs) enter the RNA-induced silencing complex and one strand guides cleavage of the target RNA by the Argonaute 2 protein1,
2,
3. A significant concern when applying siRNAs or expressing small hairpin RNAs (shRNAs) in human cells is activation of the interferon (IFN) response4,
5,
6,
7,
8,
9,
10. Synthetic siRNAs harboring certain motifs can induce an immune response when delivered to mouse and human immune cells such as peripheral blood mononuclear cells, monocytes, plasmacytoid dendritic cells (pDCs) and nonplasmacytoid dendritic cells (mDCs)11,
12,
13. In the present study we have tested the immunostimulatory effects of lipid-delivered siRNAs versus Pol III promoter–expressed shRNAs in primary CD34+ progenitor–derived hematopoietic cells. We show that in this system, lipid-delivered siRNAs are potent inducers of IFN and type I IFN gene expression, whereas the same sequences when expressed endogenously are nonimmunostimulatory.
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