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Optical In Vivo Imaging of Electrically Mediated Delivery of siRNA into Muscle for Gene Function Analysis

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Electroporation Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 423))

Abstract

Short interfering RNAs (siRNAs) represent new potential therapeutic tools owing to their capacity to induce strong, sequence-specific gene silencing in cells. However, this development requires new, safe, and efficient in vivo siRNA delivery methods. In this study, we reported that gene silencing was efficiently obtained in vivo in an adult mammal (mouse) with chemically synthesized siRNA after its electrical delivery in muscles. The associated gene silencing was followed on the same animal and lasted more than 11 days. Gene silencing was obtained in muscles not only on young adult mice but also on much older animals. No tissue damage was detected under our electrical conditions. Therefore, this method should provide an efficient approach for gene function analysis by a localized delivery of siRNAs.

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References

  1. 1. Fire, A., Xu S., Montgomery, M.K., Kostas, S.A., Driver, S.E., and Mello, C.C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 391, 806–811.

    Article  CAS  PubMed  Google Scholar 

  2. 2. McManus, M.T. and Sharp, P.A. (2002) Gene silencing in mammals by small interfering RNAs. Nat. Rev. Genet. 3, 737–747.

    Article  CAS  PubMed  Google Scholar 

  3. 3. Sioud, M. (2004) Therapeutic siRNAs. Trends Pharmacol. Sci. 25, 22–28.

    Article  CAS  PubMed  Google Scholar 

  4. 4. Zamore, P.D. and Aronin, N. (2003) siRNAs knock down hepatitis. Nat. Med. 9, 266–267.

    Article  CAS  PubMed  Google Scholar 

  5. 5. Herweijer, H. and Wolff, J.A. (2003) Progress and prospects: naked DNA gene transfer and therapy. Gene Ther. 10, 453–458.

    Article  CAS  PubMed  Google Scholar 

  6. 6. Mir, L.M., Glass, L.F., Sersa, G., et al. (1998) Effective treatment of cutaneous and subcutaneous malignant tumours by electrochemotherapy. Br. J. Cancer. 77, 2336–2342.

    Article  CAS  PubMed  Google Scholar 

  7. 7. Rols, M.P., Delteil, C., Golzio, M., Dumond, P., Cros, S., and Teissie, J. (1998) In vivo electrically mediated protein and gene transfer in murine melanoma. Nat. Biotechnol. 16, 168–171.

    Article  CAS  PubMed  Google Scholar 

  8. 8. Potts, R.O. and Chizmadzhev, Y.A. (1998) Opening doors for exogenous agents. Nat. Biotechnol. 16, 135.

    Article  CAS  PubMed  Google Scholar 

  9. 9. Heller, R., Jaraszeski, M., Atkin, A., et al. (1996) In vivo gene electroinjection and expression in rat liver. FEBS Lett. 389, 225–228.

    Article  CAS  PubMed  Google Scholar 

  10. 10. Glasspool-Malone, J., Somian, S., Drabick, J.J., and Malone, R.W. (2000) Efficient nonviral cutaneous transfection. Mol. Ther. 2, 140–146.

    Article  CAS  PubMed  Google Scholar 

  11. 11. Mathiesen, I. (1999) Electropermeabilization of skeletal muscle enhances gene transfer in vivo. Gene Ther. 6, 508–514.

    Article  CAS  PubMed  Google Scholar 

  12. 12. Aihara, H. and Miyazaki, J.-I. (1998) Gene transfer into muscle by electroporation in vivo. Nat. Biotechnol. 16, 867–870.

    Article  CAS  PubMed  Google Scholar 

  13. 13. Harrison, R.L., Byrne, B.J., and Tung, L. (1998) Electroporation-mediated gene transfer in cardiac tissue. FEBS Lett. 435, 1–5.

    Article  CAS  PubMed  Google Scholar 

  14. 14. Yang, M., Baranov, E., Li, X.M., et al. (2001) Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors. Proc. Natl. Acad. Sci. USA. 98, 2616–2621.

    Article  CAS  PubMed  Google Scholar 

  15. 15. Caplen, N.J., Parrish, S., Imani, F., Fire, A., and Morgan, R.A. (2001) Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systems. Proc. Natl. Acad. Sci. USA. 98, 9742–9747.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by grants from the CNRS CEA “Imagerie du petit animal” program, the Region Midi Pyrenees (Therapie génique et cellulaire), and the Association française contre les Myopathies.

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© 2008 Humana Press

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Golzio, M., Teissié, J. (2008). Optical In Vivo Imaging of Electrically Mediated Delivery of siRNA into Muscle for Gene Function Analysis. In: Li, S. (eds) Electroporation Protocols. Methods in Molecular Biology™, vol 423. Humana Press. https://doi.org/10.1007/978-1-59745-194-9_20

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  • DOI: https://doi.org/10.1007/978-1-59745-194-9_20

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-877-5

  • Online ISBN: 978-1-59745-194-9

  • eBook Packages: Springer Protocols

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