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PMID: 17434453 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Ectopic expression of systemic RNA interference defective protein in embryonic stem cells.

Biochemical and biophysical research communications ·Vol. 357 ·No. 2 ·2007-06-01 ·Pages 480-6

Tsang SY, Moore JC, Huizen RV, Chan CW, Li RA

Abstract

RNA interference (RNAi), a post-transcriptional gene silencing mechanism originally described in Caenorhabditis elegans, involves sequence-specific mRNA degradation mediated by double-stranded RNAs (dsRNAs). Passive dsRNA uptake has been uniquely observed in C. elegans due to the expression of systemic RNA interference defective-1 (SID-1). Here we investigated the ability of ectopic SID-1 expression to enable passive cellular uptake of short interfering RNA (siRNA) or double stranded RNA (dsRNA) in pluripotent mouse embryonic stem cells (mESCs). When SID-1-GFP and the Firefly luciferase reporter gene (luc(Fir)) were co-expressed in mESCs, luc(Fir) activity could be suppressed by simple incubation with dsRNAs/siRNAs that were designed to specifically target luc(Fir). By contrast, suppression was not observed in mESCs expressing luc(Fir) and GFP alone or when either GFP- or SID-1-GFP-expressing cells were incubated with control dsRNAs/siRNAs (non-silencing or Renilla luciferase-specific). These results may lead to high-throughput experimental strategies for studying ESC differentiation and novel approaches to genetically inhibit or eliminate the tumorigenicity of ESCs.

MeSH Terms
Animals Caenorhabditis elegans Proteins/genetics,metabolism Cell Line Embryonic Stem Cells/metabolism Gene Silencing/physiology Humans Kidney/embryology,metabolism Membrane Proteins/genetics,metabolism Mice RNA Interference/physiology Recombinant Proteins/metabolism Transfection/methods
Chemicals
Caenorhabditis elegans Proteins Membrane Proteins Recombinant Proteins SID-1 protein, C elegans
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Tsang Suk Ying
Stem Cell Program, University of California, Davis, CA 95817, USA.
Moore Jennifer C
Huizen Rika Van
Chan Camie W Y
Li Ronald A
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Article Info
Journal
Biochemical and biophysical research communications
Abbr.
Biochem Biophys Res Commun
ISSN
0006-291X
DOI
10.1016/j.bbrc.2007.03.187
Published
2007-06-01
Epub
2007-00-09
Pages
480-6
Language
English
Region
United States
NLM ID
0372516
PMCID
PMC2464293
Subset
IM
Grants
NHLBI NIH HHS · F32 HL078330 · United States
NHLBI NIH HHS · R01 HL072857 · United States
NHLBI NIH HHS · R01 HL072857-03 · United States
NHLBI NIH HHS · R01 HL72857 · United States
External Links
PubMed source DOI: 10.1016/j.bbrc.2007.03.187 PMC Full Text View journal details
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