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.

  • Paper
  • Published:

Improved packing of poly(ethylenimine)/DNA complexes increases transfection efficiency

Gene Therapy volume 6, pages 1380–1388 (1999)Cite this article

Abstract

We have developed a modified poly(ethylenimine) (PEI) transfection procedure that significantly increases PEI’s transfection efficiency. While the basic transfection procedure had a transfection efficiency of 37%, our modified procedure yielded a 53% transfection efficiency. The altered procedure gives improved results because of two simultaneous actions: free polycations are removed from the transfecting solutions, and the composition of the PEI complexes that are administered to cells has been modified. The reduction in the amount of free polycations in transfecting solutions reduced the toxicity sometimes associated with the administration of polycations to cellular environments. The structural modification of PEI/DNA transfecting complexes involves improved PEI packing around the delivered plasmid to yield a greater buffering capacity without a change in the complex’s surface charge concentration. These structural properties were confirmed by titration and ζ potential analyses. Whether the modified PEI/DNA complexes are more effective because of increased cellular uptake or an enhanced ability to escape from endolysosomes has been addressed. The increase in transfection efficiency was obtained when the buffering capacity of the PEI/DNA was increased without a change in surface charge concentration, which implies that it is the property of enhanced lysosomal buffering that is responsible for successful PEI transfection.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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. Boussif O et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine Proc Natl Acad Sci USA 1995 92: 7297–7301

    Article  CAS  Google Scholar 

  2. Lambert RC et al. Polyethylenimine-mediated DNA transfection of peripheral and central neurons in primary culture: probing Ca2+ channel structure and function with antisense oligonucleotides Molec and Cell Neurosci 1996 7: 239–246

    Article  CAS  Google Scholar 

  3. Boussif O, Zanta MA, Behr JP . Optimized galenics improve in vitro gene transfer with cationic molecules up to 1000-fold Gene Therapy 1996 3: 1074–1080

    CAS  Google Scholar 

  4. Abdallah B et al. A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: polyethylenimine Hum Gene Ther 1996 7: 1947–1954

    Article  CAS  Google Scholar 

  5. Boletta A et al. Nonviral gene delivery to the rat kidney with polyethylenimine Hum Gene Ther 1997 8: 1243–1251

    Article  CAS  Google Scholar 

  6. Godbey WT, Wu KK, Mikos AG . Size matters: molecular weight affects the efficiency of poly(ethylenimine) as a gene delivery vehicle J Biomed Mat Res 1999 45: 268–275

    Article  CAS  Google Scholar 

  7. Dunlap DD, Maggi AS, Marco R, Monaco L . Nanoscopic structure of DNA condensed for gene delivery Nucleic Acids Res 1997 25: 3095–3101

    Article  CAS  Google Scholar 

  8. Helander IM, Latva-Kala K, Lounatmaa K . Permeabilizing action of polyethylenimine on Salmonella typhimurium involves disruption of the outer membrane with lipopolysaccharide Microbiology 1998 144: 385–390

    Article  CAS  Google Scholar 

  9. Widom J, Baldwin RL . Cation-induced toroidal condensation of DNA studies with Co3+(NH3)6 J Molec Biol 1980 144: 431–453

    Article  CAS  Google Scholar 

  10. Kim JS, Maruyama A, Akaike T, Kim SW . In vitro gene expression on smooth muscle cells using a terplex delivery system J Control Rel 1997 47: 51–59

    Article  CAS  Google Scholar 

  11. Labat-Moleur F et al. An electron microscopy study into the mechanism of gene transfer with lipopolyamines Gene Therapy 1996 3: 1010–1017

    CAS  Google Scholar 

  12. Nemati F . PhD. dissertation University of Paris IX 1996

    Google Scholar 

  13. Helander IM, Alakomi HL, Latva-Kala K, Koski P . Polyethylenimine is an effective permeabilizer of gram-negative bacteria Microbiology 1997 143: 3193–3199

    Article  CAS  Google Scholar 

  14. Godbey WT, Wu KK, Mikos AG . Tracking the intracellular path of poly(ethylenimine) for gene delivery Proc Natl Acad Sci USA 1999 96: 5177–5181

    Article  CAS  Google Scholar 

  15. Ferrari S et al. ExGen 500 is an efficient vector for gene delivery to lung epithelial cells in vitro and in vivo Gene Therapy 1997 4: 1100–1106

    Article  CAS  Google Scholar 

  16. Plank C, Mechtler K, Szoka FC, Wagner E . Activation of the complement system by synthetic DNA complexes: a potential barrier for intravenous gene delivery Hum Gene Ther 1996 7: 1437–1446

    Article  CAS  Google Scholar 

  17. Hunter RJ . Zeta Potential in Colloid Science: Principles and Applications Academic Press: New York 1981

    Google Scholar 

  18. Edgell CJ, McDonald C, Graham JB . Permanent cell line expressing human factor VIII-related antigen established by hybridization Proc Natl Acad Sci USA 1983 80: 3734–3737

    Article  CAS  Google Scholar 

  19. Suggs JE, Madden MC, Friedman M, Edgell CJ . Prostacyclin expression by a continuous human cell line derived from vascular endothelium Blood 1986 68: 825–829

    CAS  PubMed  Google Scholar 

  20. Bailey FE, Koleske JV . Solution properties of polyl(ethylene oxide) Poly(Ethylene Oxide) Academic Press: New York 1976 pp 29–86

    Chapter  Google Scholar 

Download references

Acknowledgements

This material is based upon work supported under a National Science Foundation Graduate Fellowship (WTG) and the National Institutes of Health (R29-AR42639) (AGM), (PSO-NS-23327) (KKW), and (R01-HL-50675) (KKW). We also wish to thank Nancy Turner and Dr Tom Chow for their assistance with the FACScan, as well as Steve Yang for his guidance in the use of the DELSA 440 (ζ potential). We are also grateful to Dr Cora-Jean Edgell for providing the EA.hy 926 cell line.

Author information

Authors and Affiliations

  1. Department of Biochemistry and Cell Biology, Rice University, Houston, USA

    W T Godbey

  2. Division of Hematology and Vascular Biology Research Center, The University of Texas Health Science Center at Houston, Houston, USA

    K K Wu

  3. Department of Chemical Engineering, Rice University, Houston, TX, USA

    G J Hirasaki & A G Mikos

  4. Department of Bioengineering, Rice University, Houston, TX, USA

    A G Mikos

Authors
  1. W T Godbey
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. K K Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G J Hirasaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A G Mikos
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Godbey, W., Wu, K., Hirasaki, G. et al. Improved packing of poly(ethylenimine)/DNA complexes increases transfection efficiency. Gene Ther 6, 1380–1388 (1999). https://doi.org/10.1038/sj.gt.3300976

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3300976

Keywords

This article is cited by

Search

Advanced search

Quick links