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Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation
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Chapters
- 00:05Title
- 01:05Program the Amaxa 96-well Shuttle Nucleofactor
- 02:00Prepare DCs for Transfection
- 03:40Transfect DCs
- 05:07Infection and Harvest of Cells with Newcastle Disease Virus
- 06:09siRNA Gene Targeting Results in Drastic Reduction of Message and Total Reduction of Protein Expression in Monocyte DCs
- 07:51Conclusion
We present our optimized high-throughput nucleofection protocol as an efficient way of transfecting primary human monocyte-derived dendritic cells with either plasmid DNA or siRNA without causing cell maturation. We further provide evidence for successful siRNA silencing of targeted gene RIG-I at both the mRNA and protein levels.
Tags
Dendritic CellsImmune SystemInfectionPathogen-associated Molecular Patterns (PAMPs)Pattern Recognition Receptors (PRRs)Cell MaturationType 1 InterferonProinflammatory CytokinesMHCIICD86Adaptive Immune ResponseMolecular NetworksSignaling PathwaysInduced GenesDC-based VaccinesTransfection Of Dendritic Cells
