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Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation
Published on: October 20, 2014
Ectopic expression of systemic RNA interference defective protein in embryonic stem cells
Suk Ying Tsang1, Jennifer C Moore, Rika Van Huizen
1Stem Cell Program, University of California, Davis, CA 95817, USA.
Biochemical and Biophysical Research Communications
|April 17, 2007
Summary
Mouse embryonic stem cells (mESCs) can take up double-stranded RNA (dsRNA) when expressing the SID-1 gene. This enables sequence-specific gene silencing, offering new research and therapeutic strategies for stem cells.
Area of Science:
- Molecular Biology
- Genetics
- Stem Cell Biology
Background:
- RNA interference (RNAi) is a gene silencing mechanism.
- Passive dsRNA uptake is unique to C. elegans via SID-1.
- Mouse embryonic stem cells (mESCs) lack natural dsRNA uptake mechanisms.
Purpose of the Study:
- To investigate if ectopic SID-1 expression enables passive dsRNA/siRNA uptake in mESCs.
- To explore the potential for SID-1 mediated gene silencing in mESCs.
Main Methods:
- Co-expression of SID-1-GFP and Firefly luciferase reporter in mESCs.
- Incubation of mESCs with specific dsRNAs/siRNAs targeting luciferase.
- Control experiments with GFP-expressing cells and non-targeting dsRNAs/siRNAs.
Main Results:
- Ectopic SID-1 expression enabled passive cellular uptake of dsRNA/siRNA in mESCs.
- Incubation with specific dsRNAs/siRNAs suppressed luciferase activity in SID-1 expressing mESCs.
- No suppression was observed in control mESCs or with non-specific dsRNAs/siRNAs.
Conclusions:
- SID-1 expression confers the ability for passive dsRNA/siRNA uptake and gene silencing in mESCs.
- This finding opens avenues for high-throughput screening in ESC differentiation.
- Potential applications include novel strategies to control ESC tumorigenicity.
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