Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Non-viral gene transfer of murine spleen cells achieved by in vivo electroporation

Abstract

Gene electrotranfer is an attractive physical method to deliver genes to target tissues. The aim of this study was to evaluate in vivo gene electrotransfer into spleen, one of the most important lymphoid organ, in order to create a new tool to modulate the immuno-inflammatory system. C57Bl/6 mice were submitted either to intramuscular electrotransfer (IME) as a reference method or to intrasplenic (ISE) gene electrotransfer. In the naked injected plasmids, the CMV promoter controlled the expression of luciferase, secreted alkaline phosphatase, EGFP, or IFNγ. The ISE optimal electrotransfer conditions were first determined and ISE was found to be an efficient gene transfer method, which can be used to express secreted or intracellular proteins transiently. Although transfected cells were still present in the spleen 30 days after ISE, transfected spleen cells could recirculate since they were detected in extrasplenic locations. Using a T-lymphocyte-specific promoter controlling the expression of EGFP, splenic T cells could be targeted. Finally, it appeared that ISE procedure does not impair by itself the immune response and does not result in a significant production of antibodies directed to the transgenic proteins in C57Bl/6 mice. This strategy constitutes a new method to manipulate the immune response that can be used in various experimental designs.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  1. Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH . Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J 1982; 1: 841–845.

    Article  CAS  Google Scholar 

  2. Klenchin VA et al. Electrically induced DNA uptake by cells is a fast process involving DNA electrophoresis. Biophys J 1991; 60: 804–811.

    Article  CAS  Google Scholar 

  3. Zheng QA, Chang DC . High-efficiency gene transfection by in situ electroporation of cultured cells. Biochim Biophys Acta 1991; 1088: 104–110.

    Article  CAS  Google Scholar 

  4. Sukharev SI et al. Electroporation and electrophoretic DNA transfer into cells. The effect of DNA interaction with electropores. Biophys J 1992; 63: 1320–1327.

    Article  CAS  Google Scholar 

  5. Tripathy SK et al. Stable delivery of physiologic levels of recombinant erythropoietin to the systemic circulation by intramuscular injection of replication- defective adenovirus. Proc Natl Acad Sci USA 1994; 91: 11557–11561.

    Article  CAS  Google Scholar 

  6. Tsurumi Y et al. Direct intramuscular gene transfer of naked DNA encoding vascular endothelial growth factor augments collateral development and tissue perfusion. Circulation 1996; 94: 3281–3290.

    Article  CAS  Google Scholar 

  7. Naffakh N et al. Long-term secretion of therapeutic proteins from genetically modified skeletal muscles. Hum Gene Ther 1996; 7: 11–21.

    Article  CAS  Google Scholar 

  8. Miller G et al. Expression of factor VII by muscle cells in vitro and in vivo following direct gene transfer: modelling gene therapy for haemophilia. Gene Ther 1995; 2: 736–742.

    CAS  PubMed  Google Scholar 

  9. Rizzuto G et al. Efficient and regulated erythropoietin production by naked DNA injection and muscle electroporation. Proc Natl Acad Sci USA 1999; 96: 6417–6422.

    Article  CAS  Google Scholar 

  10. Piccirillo CA, Prud'homme GJ . Prevention of experimental allergic encephalomyelitis by intramuscular gene transfer with cytokine-encoding plasmid vectors. Hum Gene Ther 1999; 10: 1915–1922.

    Article  CAS  Google Scholar 

  11. Schratzberger P et al. Reversal of experimental diabetic neuropathy by VEGF gene transfer. J Clin Invest 2001; 107: 1083–1092.

    Article  CAS  Google Scholar 

  12. Montgomery DL, Ulmer JB, Donnelly JJ, Liu MA . DNA vaccines. Pharmacol Ther 1997; 74: 195–205.

    CAS  PubMed  Google Scholar 

  13. Titomirov AV, Sukharev S, Kistanova E . In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA. Biochim Biophys Acta 1991; 1088: 131–134.

    Article  CAS  Google Scholar 

  14. Heller R et al. In vivo gene electroinjection and expression in rat liver. FEBS Lett 1996; 389: 225–228.

    Article  CAS  Google Scholar 

  15. Suzuki T et al. Direct gene transfer into rat liver cells by in vivo electroporation. FEBS Lett 1998; 425: 436–440.

    Article  CAS  Google Scholar 

  16. Rols MP et al. In vivo electrically mediated protein and gene transfer in murine melanoma. Nat Biotechnol 1998; 16: 168–171.

    Article  CAS  Google Scholar 

  17. Aihara H, Miyazaki J . Gene transfer into muscle by electroporation in vivo. Nat Biotechnol 1998; 16: 867–870.

    Article  CAS  Google Scholar 

  18. Mir LM et al. Long-term, high level in vivo gene expression after electric pulse-mediated gene transfer into skeletal muscle. CR Acad Sci HI 1998; 321: 893–899.

    Article  CAS  Google Scholar 

  19. Mir LM et al. High-efficiency gene transfer into skeletal muscle mediated by electric pulses. Proc Natl Acad Sci USA 1999; 96: 4262–4267.

    Article  CAS  Google Scholar 

  20. Bettan M, Darteil R, Scherman D . Secreted human placental alkaline phosphatase as a reporter gene for in vivo gene transfer. Anal Biochem 1999; 271: 187–189.

    Article  CAS  Google Scholar 

  21. Kreiss P, Bettan M, Crouzet J, Scherman D . Erythropoietin secretion and physiological effect in mouse after intramuscular plasmid DNA electrotransfer. J Gene Med 1999; 1: 245–250.

    Article  CAS  Google Scholar 

  22. Bettan M et al. High-level protein secretion into blood circulation after electric pulse-mediated gene transfer into skeletal muscle. Mol Ther 2000; 2: 204–210.

    Article  CAS  Google Scholar 

  23. Payen E et al. Improvement of mouse beta-thalassemia by electrotransfer of erythropoietin cDNA. Exp Hematol 2001; 29: 295–300.

    Article  CAS  Google Scholar 

  24. Contag CH et al. Photonic detection of bacterial pathogens in living hosts. Mol Microbiol 1995; 18: 593–603.

    Article  CAS  Google Scholar 

  25. Contag CH et al. Visualizing gene expression in living mammals using a bioluminescent reporter. Photochem Photobiol 1997; 66: 523–531.

    Article  CAS  Google Scholar 

  26. Contag PR, Olomu IN, Stevenson DK, Contag CH . Bioluminescent indicators in living mammals. Nat Med 1998; 4: 245–247.

    Article  CAS  Google Scholar 

  27. Mennuni C et al. Hyaluronidase increases electrogene transfer efficiency in skeletal muscle. Hum Gene Ther 2002; 13: 355–365.

    Article  CAS  Google Scholar 

  28. Salmon P et al. Characterization of an intronless CD4 minigene expressed in mature CD4 and CDS T cells, but not expressed in immature thymocytes. J Immunol 1996; 156: 1873–1879.

    CAS  Google Scholar 

  29. Velikovsky CA et al. Single-shot plasmid DNA intrasplenic immunization for the production of monoclonal antibodies. Persistent expression of DNA. J Immunol Methods 2000; 244: 1–7.

    Article  CAS  Google Scholar 

  30. Soubrier F et al. pCOR: a new design of plasmid vectors for nonviral gene therapy. Gene Ther 1999; 6: 1482–1488.

    Article  CAS  Google Scholar 

  31. Nicoletti A et al. Immunoglobulin treatment reduces atherosclerosis in apo E knockout mice. J Clin Invest. 1998; 102: 910–918.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by INSERM and by a grant from the Association Claude Bernard. B Poirier was the recipient of a fellowship from GRRC and E Tupin was the recipient of a fellowship from the ARCOL.

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tupin, E., Poirier, B., Bureau, M. et al. Non-viral gene transfer of murine spleen cells achieved by in vivo electroporation. Gene Ther 10, 569–579 (2003). https://doi.org/10.1038/sj.gt.3301914

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301914

Keywords

This article is cited by

Search

Quick links