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Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation
JoVE 신문
면역학 및 감염병학
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JoVE 신문 면역학 및 감염병학
Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation
DOI:

08:08 min

July 08, 2011

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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

Summary

자동 번역

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.

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