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:

Cationic lipid-mediated transfection of cells in culture requires mitotic activity

Gene Therapy volume 6, pages 403–411 (1999)Cite this article

Abstract

Cationic lipid-based delivery systems such as lipoplexes or stabilized plasmid–lipid particles (SPLP) represent a safer alternative to viral systems for gene therapy applications. We studied the impact of cell cycle status on the efficiency of transfection of human ovarian carcinoma tumor cells using two cationic-lipid based delivery systems. Cells arrested in the G1 phase of the cell cycle by treatment with aphidicolin were compared with an asynchronous dividing population of cells. Treatment of the cells with aphidicolin had no effect on the rate of internalization of the lipid formulated DNA or on the level of gene expression observable in stably transfected cells. However, cells treated with aphidicolin exhibited 20-fold lower reporter gene activity than asynchronous control cells upon incubation with lipoplexes. When cells arrested in the G1 phase were allowed to proceed though the cell cycle in the presence of the lipoplex or SPLP, transgene expression was found to coincide with the transition of cells from the G2/M phase into the G1 phase of the subsequent cell cycle. In addition, higher levels of reporter gene expression were observed when the cells were incubated with lipoplexes or SPLP during, or just before, mitosis. These results suggest that it may be possible to augment cationic lipid-mediated transfection by manipulating the cell cycle status of the target cells.

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

Similar content being viewed by others

References

  1. Felgner PL et al. Lipofectin: a highly efficient lipid-mediated DNA transfection procedure Proc Natl Acad Sci USA 1987 84: 7413–7417

    Article  CAS  Google Scholar 

  2. Felgner PL et al. Improved cationic lipid formulations for in vivo gene therapy Ann NY Acad Sci 1995 772: 126–139

    Article  CAS  Google Scholar 

  3. Gao X, Huang L . A novel cationic liposome reagent for efficient transfection of mammalian cells Biochem Biophys Res Commun 1991 179: 280–285

    Article  CAS  Google Scholar 

  4. Caplen NJ et al. Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis Nature Med 1995 1: 39–46

    Article  CAS  Google Scholar 

  5. Nabel GJ et al. Immune response in human melanoma after transfection of an allogenic class I major histocompatibility complex gene with DNA liposome Proc Natl Acad Sci USA 1996 93: 15388–15399

    Article  CAS  Google Scholar 

  6. Gill DR et al. A placebo controlled study of liposome-mediated gene transfer to the nasal epithelium of patients with cystic fibrosis Gene Therapy 1997 4: 199–209

    Article  CAS  Google Scholar 

  7. Zabner J et al. Cellular and molecular barriers to gene transfer by a cationic lipid J Biol Chem 1995 270: 18997–19007

    Article  CAS  Google Scholar 

  8. Wrobel I, Collins D . Fusion of cationic liposomes with mammalian cells occurs after endocytosis Biochim Biophys Acta 1995 1235: 296–304

    Article  Google Scholar 

  9. Xu Y, Szoka FC Jr . Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection Biochemistry 1996 35: 5616–5623

    Article  CAS  Google Scholar 

  10. Friend DS, Papahadjopoulos D, Debs RJ . Endocytosis and intracellular processing accompanying transfection mediated by cationic liposomes Biochim Biophys Acta 1996 1278: 41–50

    Article  Google Scholar 

  11. Wilke M et al. Efficacy of a peptide-based gene delivery system depends on mitotic activity Gene Therapy 1996 3: 1133–1142

    CAS  PubMed  Google Scholar 

  12. Nicolau C, Sene C . Liposome-mediated DNA transfer in eukaryotic cells Biochim Biophys Acta 1982 721: 185–190

    Article  CAS  Google Scholar 

  13. Fasbender A, Zabner J, Zeiher BG, Welsh MJ . A low rate of cell proliferation and reduced DNA uptake limit cationic lipid-mediated gene transfer to primary cultures of ciliated human airway epithelia Gene Therapy 1997 4: 1173–1180

    Article  CAS  Google Scholar 

  14. Jiang C et al. Efficiency of cationic lipid-mediated transfection of polarized and differentiated airway epithelial cells in vitro and in vivo Hum Gene Ther 1998 9: 1531–1542

    Article  CAS  Google Scholar 

  15. Bukrinsky MI et al. A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells Nature 1993 365: 666–669

    Article  CAS  Google Scholar 

  16. Naldini L et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector Science 1996 272: 263–267

    Article  CAS  Google Scholar 

  17. von Schwedler U, Kornbluth RS, Trono D . The nuclear localization signal of the matrix protein of human immunodeficiency virus type 1 allows the establishment of infection in macrophages and quiescent T lymphocytes Proc Natl Acad Sci USA 1994 91: 6992–6996

    Article  CAS  Google Scholar 

  18. Felgner PL et al. Nomenclature for synthetic gene delivery systems Hum Gene Ther 1997 8: 511–512

    Article  CAS  Google Scholar 

  19. Wheeler J et al. Stabilized plasmid-lipid particles: construction and characterization Gene Therapy 1999 (in press)

  20. Ikegami S et al. Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-alpha Nature 1978 275: 458–460

    Article  CAS  Google Scholar 

  21. Wahl AF et al. Properties of two forms of DNA polymerase delta from calf thymus Biochemistry 1986 25: 7821–7827

    Article  CAS  Google Scholar 

  22. Tseng W-C et al. Cationic liposomal delivery of plasmid to endothelial cells measured by quantitative flow cytometry Biotechnol Bioeng 1996 50: 548–554

    Article  CAS  Google Scholar 

  23. Tseng W-C, Haselton FR, Giorgio TD . Transfection of cationic liposomes using simultaneous single cell measurements of plasmid delivery and transgene expression J Biol Chem 1997 272: 25641–25647

    Article  CAS  Google Scholar 

  24. Mulligan RC . The basic science of gene therapy Science 1993 260: 926–932

    Article  CAS  Google Scholar 

  25. Verma IM . Gene therapy: hopes, hypes, and hurdles Molec Med 1994 1: 2–3

    Article  CAS  Google Scholar 

  26. Greber UF et al. Stepwise dismantling of adenovirus 2 during entry into cells Cell 1993 75: 477–486

    Article  CAS  Google Scholar 

  27. Dowty J et al. Plasmid DNA entry into postmitotic nuclei of primary rat myotubules Proc Natl Acad Sci USA 1995 92: 4572–4576

    Article  CAS  Google Scholar 

  28. Maniatis T, Fritsch EF, Sambrook J . Molecular Cloning Cold Spring Harbor: New York, 1981, pp 93–94

  29. Liljestrom P, Garoff H . Expression of proteins using Semliki forest virus vectors. In: Ausubel FM et al (eds). Current Protocols in Molecular Biology John Wiley: Canada 1994 (Suppl. 28) 16.20.1–16.20.16

    Google Scholar 

  30. Lechardeur D et al. Metabolic instability of plasmid DNA in the cytosol; a potential barrier to gene transfer Gene Therapy 1999 (in press)

Download references

Author information

Authors and Affiliations

  1. Inex Pharmaceuticals Corporation, Burnaby, British Columbia, Canada

    I Mortimer, P Tam, I MacLachlan, R W Graham, E G Saravolac & P B Joshi

Authors
  1. I Mortimer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P Tam
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. I MacLachlan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R W Graham
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E G Saravolac
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. P B Joshi
    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

Mortimer, I., Tam, P., MacLachlan, I. et al. Cationic lipid-mediated transfection of cells in culture requires mitotic activity. Gene Ther 6, 403–411 (1999). https://doi.org/10.1038/sj.gt.3300837

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3300837

Keywords

This article is cited by

Search

Advanced search

Quick links