TY - JOUR
T1 - Stable expression of shRNAs in human CD34+ progenitor cells can avoid induction of interferon responses to siRNAs in vitro
AU - Robbins, Marjorie A.
AU - Li, Mingjie
AU - Leung, Irene
AU - Li, Haitang
AU - Boyer, Doris V.
AU - Song, Yong
AU - Behlke, Mark A.
AU - Rossi, John J.
N1 - Funding Information:
The authors would like to thank Lucy Brown and Claude Yosballa at the COH FACS Facility for FACS assays and analysis. This work was supported by National Institutes of Health grants AI29329, AI42552, HL07470 and AI061389.
PY - 2006/5
Y1 - 2006/5
N2 - 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 protein1-3. A significant concern when applying siRNAs or expressing small hairpin RNAs (shRNAs) in human cells is activation of the Interferon (IFN) response4-10. 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)11-13. 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 IFNα and type I IFN gene expression, whereas the same sequences when expressed endogenously are nonimmunostimulatory.
AB - 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 protein1-3. A significant concern when applying siRNAs or expressing small hairpin RNAs (shRNAs) in human cells is activation of the Interferon (IFN) response4-10. 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)11-13. 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 IFNα and type I IFN gene expression, whereas the same sequences when expressed endogenously are nonimmunostimulatory.
UR - http://www.scopus.com/inward/record.url?scp=33646588689&partnerID=8YFLogxK
U2 - 10.1038/nbt1206
DO - 10.1038/nbt1206
M3 - Article
C2 - 16648841
AN - SCOPUS:33646588689
SN - 1087-0156
VL - 24
SP - 566
EP - 571
JO - Nature Biotechnology
JF - Nature Biotechnology
IS - 5
ER -