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Long-term Silencing of Intersectin-1s in Mouse Lungs by Repeated Delivery of a Specific siRNA via Cationic Liposomes. Evaluation of Knockdown Effects by Electron Microscopy
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Biologia
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JoVE Journal Biologia
Long-term Silencing of Intersectin-1s in Mouse Lungs by Repeated Delivery of a Specific siRNA via Cationic Liposomes. Evaluation of Knockdown Effects by Electron Microscopy

Long-term Silencing of Intersectin-1s in Mouse Lungs by Repeated Delivery of a Specific siRNA via Cationic Liposomes. Evaluation of Knockdown Effects by Electron Microscopy

DOI:
10.3791/50316-v

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15:55 min

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June 21, 2013

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, ,
1Department of Pharmacology,Rush University, 2Pulmonary and Critical Care Department,Rush University

Capitoli

  • 00:05Titolo
  • 02:06Prepare Cationic Liposomes and Liposomes:siRNA-ITSN-1 Complexes
  • 05:30Mouse Retro-orbital Vein Injection (Internal Angle of the Eye Socket)
  • 06:39Mouse Lung Perfusion and Tissue Collection for Biochemical Studies
  • 08:23Mouse Lung Tissue Processing for EM
  • 10:21Results: Down-regulation of Intersectin-1 Expression Affects Morphology of Lung Tissue
  • 15:14Conclusion

Summary

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Traduzione automatica

Repeated retro-orbital injections of cationic liposomes/siRNA/ITSN-1s complexes in mice, every 72 hours for 24 days, efficiently deliver the siRNA duplex to the mouse lung microvasculature reducing ITSN-1s mRNA and protein expression by 75%. This technique is highly reproducible in an animal model, has no adverse effects and avoids fatalities.

Tags

Keywords: SiRNAITSN-1sCationic LiposomesLung EndotheliumElectron MicroscopyGene SilencingMouse ModelVascular PermeabilityEndothelial Cell Survival

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