Ectopic expression of systemic RNA interference defective protein in embryonic stem cells
| Autor: | Jennifer C. Moore, Suk Ying Tsang, Rika Van Huizen, Ronald A. Li, Camie W.Y. Chan |
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| Rok vydání: | 2007 |
| Předmět: |
Embryonic stem cells
Small interfering RNA Biophysics SID-1 Caenorhabditis elegans Proteins - genetics - metabolism Biology Kidney Transfection Biochemistry Article Cell Line Mice RNA interference Membrane Proteins - genetics - metabolism Animals Humans Gene silencing Gene Silencing Caenorhabditis elegans Proteins Gene transfer Kidney - embryology - metabolism Molecular Biology Reporter gene Membrane Proteins Kidney metabolism Cell Biology Embryonic stem cell Molecular biology Recombinant Proteins RNA silencing Ectopic expression Gene Silencing - physiology Embryonic Stem Cells - metabolism |
| Zdroj: | Biochemical and Biophysical Research Communications. 357:480-486 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2007.03.187 |
| Popis: | 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 (lucFir) were co-expressed in mESCs, lucFir activity could be suppressed by simple incubation with dsRNAs/siRNAs that were designed to specifically target lucFir. By contrast, suppression was not observed in mESCs expressing lucFir 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. © 2007 Elsevier Inc. All rights reserved. link_to_OA_fulltext |
| Databáze: | OpenAIRE |
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