Skip to main content

Advertisement

SpringerLink
Log in

Polyethylenimine-Based Transfection Method as a Simple and Effective Way to Produce Recombinant Lentiviral Vectors

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

HIV-1-derived lentiviral vectors (LvV) are within the most attractive gene delivery vehicles in the context of both dividing and quiescent cells. LvV is currently produced by the conventional calcium phosphate precipitation method. Nevertheless, this procedure is highly susceptible to variations in pH and impurities, which lead to inconsistencies in LvV production. Here, we present a simple and robust procedure for LvV production using branched 25 kDa polyethylenimine, with a transfection efficiency of over 90% and viral titer yields of about 1 × 107 infective lentiviral particles per milliliter. The procedure outlined is simple, consistent, and as inexpensive as the CaPO4-based method.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Iwakuma, T., Cui, Y., & Chang, L. J. (1999). Virology, 261, 120–132. doi:10.1006/viro.1999.9850.

    Article  CAS  Google Scholar 

  2. Miyoshi, H., Blomer, U., Takahashi, M., Gage, F. H., & Verma, I. M. (1998). Journal of Virology, 72, 8150–8157.

    CAS  Google Scholar 

  3. De Palma, M., & Naldini, L. (2002). Methods in Enzymology, 346, 514–529. doi:10.1016/S0076-6879(02)46074-0.

    Article  Google Scholar 

  4. Yu, H., Rabson, A. B., Kaul, M., Ron, Y., & Dougherty, J. P. (1996). Journal of Virology, 70, 4530–4537.

    CAS  Google Scholar 

  5. Kaul, M., Yu, H., Ron, Y., & Dougherty, J. P. (1998). Virology, 249, 167–174. doi:10.1006/viro.1998.9327.

    Article  CAS  Google Scholar 

  6. Rohll, J. B., Mitrophanous, K. A., Martin-Rendon, E., Ellard, F. M., Radcliffe, P. A., Mazarakis, N. D., et al. (2002). Methods in Enzymology, 346, 466–500. doi:10.1016/S0076-6879(02)46072-7.

    Article  CAS  Google Scholar 

  7. Naldini, L., Blomer, U., Gallay, P., Ory, D., Mulligan, R., Gage, F. H., et al. (1996). Science, 272, 263–267. doi:10.1126/science.272.5259.263.

    Article  CAS  Google Scholar 

  8. Coleman, J. E., Huentelman, M. J., Kasparov, S., Metcalfe, B. L., Paton, J. F., Katovich, M. J., et al. (2003). Physiological Genomics, 12, 221–228.

    CAS  Google Scholar 

  9. Shin, K. J., Wall, E. A., Zavzavadjian, J. R., Santat, L. A., Liu, J., Hwang, J. I., et al. (2006). Proceedings of the National Academy of Sciences of the United States of America, 103, 13759–13764. doi:10.1073/pnas.0606179103.

    Article  CAS  Google Scholar 

  10. Kosaka, Y., Kobayashi, N., Fukazawa, T., Totsugawa, T., Maruyama, M., Yong, C., et al. (2004). Artificial Organs, 28, 271–277. doi:10.1111/j.1525-1594.2004.47297.x.

    Article  CAS  Google Scholar 

  11. Segura, M. M., Garnier, A., Durocher, Y., Coelho, H., & Kamen, A. (2007). Biotechnology and Bioengineering, 98, 789–799. doi:10.1002/bit.21467.

    Article  CAS  Google Scholar 

  12. Emi, N., Friedmann, T., & Yee, J. K. (1991). Journal of Virology, 65, 1202–1207.

    CAS  Google Scholar 

  13. Abdallah, B., Hassan, A., Benoist, C., Goula, D., Behr, J. P., & Demeneix, B. A. (1996). Human Gene Therapy, 7, 1947–1954. doi:10.1089/hum.1996.7.16-1947.

    Article  CAS  Google Scholar 

  14. Tiscornia, G., Singer, O., & Verma, I. M. (2006). Nature Protocols, 1, 241–245. doi:10.1038/nprot.2006.37.

    Article  CAS  Google Scholar 

  15. Huang, M. B., Khan, M., Garcia-Barrio, M., Powell, M., & Bond, V. C. (2001). J Acq Immun Def Synd, 27, 213–221.

    CAS  Google Scholar 

Download references

Acknowledgments

We thank Dr. Jose Alberto Cremata and Dr. Yamila Carpio for their critical comments and fruitful discussions on the manuscript.

Author information

Authors and Affiliations

  1. Animal Biotechnology Department, Centre for Genetic Engineering and Biotechnology, Havana, Cuba

    Jorge R. Toledo, Yanet Prieto, Nayrobis Oramas & Oliberto Sánchez

  2. Department of Transgenesis and Animal Cloning, Animal Biotechnology Division, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana, 10600, Cuba

    Oliberto Sánchez

Authors
  1. Jorge R. Toledo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yanet Prieto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nayrobis Oramas
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Oliberto Sánchez
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Oliberto Sánchez.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Toledo, J.R., Prieto, Y., Oramas, N. et al. Polyethylenimine-Based Transfection Method as a Simple and Effective Way to Produce Recombinant Lentiviral Vectors. Appl Biochem Biotechnol 157, 538–544 (2009). https://doi.org/10.1007/s12010-008-8381-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-008-8381-2

Keywords

Search

Navigation