Efficient In Vivo Delivery of Plasmids and Oligonucleotides Using Hemagglutinating Virus of Japan Envelope (HVJ‐E) Vector in Immunological Disease Models

Atsushi Kitani1, Stefan Fichtner‐Feigl2

1 National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, 2 University of Regensburg, Regensburg, Germany
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 10.17E
DOI:  10.1002/0471142735.im1017es91
Online Posting Date:  November, 2010
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This unit describes a method for in vivo delivery of oligonucleotides or plasmids using the hemagglutinating virus of Japan envelope (HVJ‐E), an inactivated Sendai virus particle, as a delivery system. Viral transfection methods generally show a higher transfection efficiency than nonviral methods for the delivery of genes to cells. However, in using these methods one must bear in mind that the introduction of a virus particle into a host carries a risk for leukemia induction and for creation of disturbances in immune function due to cytotoxicity. Curr. Protoc. Immunol. 91:10.17E.1‐10.17E.9. © 2010 by John Wiley & Sons, Inc.

Keywords: hemagglutinating virus of Japan; Sendai virus; plasmid; siRNA; decoy oligo; gene therapy; in vivo

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Table of Contents

  • Introduction
  • Basic Protocol 1: Plasmid Packaging into the HVJ‐E Particle
  • Alternate Protocol 1: Oligonucleotides Delivery with HVJ‐E Particle
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
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Basic Protocol 1: Plasmid Packaging into the HVJ‐E Particle

  • HVJ envelope (see recipe), ice‐cold
  • 1 mg/ml protamine sulfate/PBS (see recipe), ice‐cold
  • Plasmid
  • Dulbecco's phosphate‐buffered saline (D‐PBS)
  • 3% Triton X‐100 (see recipe), ice‐cold
  • 1.7‐ml microcentrifuge tubes
  • 1‐ml syringes equipped with 26‐G needles
NOTE: All equivalent materials described above, other than the plasmid, are contained in the GenomONE Neo EX HVJ Envelope Transfection Kit supplied from Ishihara Sangyo Kaisha or Cosmo Bio (http://www.cosmobio.co.jp/export_e/products/cells/products_ISK_20070531.asp).

Alternate Protocol 1: Oligonucleotides Delivery with HVJ‐E Particle

  • Oligonucleotides (siSTABLE siRNA from Dharmacon/Thermoscientific or phosphothioate DNA) dissolved at the concentration of 10 mg/ml in nuclease free, ice‐cold PBS
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Literature Cited

Literature Cited
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   Fichtner‐Feigl, S., Fuss, I.J., Preiss, J.C., Strober, W., and Kitani, A. 2005. Treatment of murine Th1‐ and Th2‐mediated inflammatory bowel disease with NF‐kappa B decoy oligonucleotides. J. Clin. Invest. 115:3057‐3071.
   Fichtner‐Feigl, S., Strober, W., Kawakami, K., Puri, R.K., and Kitani, A. 2006. IL‐13 signaling through the IL‐13alpha2 receptor is involved in induction of TGF‐beta1 production and fibrosis. Nat. Med. 12:99‐106.
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