title: Ectopic expression of systemic RNA interference defective protein in embryonic stem cells.
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.
authors: Stem Cell Program, University of California, Davis, CA 95817, USA.
authors: Suk Ying
authors: SY
authors: Tsang
authors: Jennifer C
authors: JC
authors: Moore
authors: Rika Van
authors: RV
authors: Huizen
authors: Camie W Y
authors: CW
authors: Chan
authors: Ronald A
authors: RA
authors: Li
mesh_terms: D000818:Animals; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000378:metabolism*; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D007668:Kidney / Q000196:embryology* / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D034622:RNA Interference / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D029742:Caenorhabditis elegans Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; C452203:SID-1 protein, C elegans