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.

  • Brief Communication
  • Published:

Muscle-specific enhancement of gene expression by incorporation of SV40 enhancer in the expression plasmid

Gene Therapy volume 8, pages 494–497 (2001)Cite this article

Abstract

Skeletal muscle is established as an ideal tissue for gene delivery to treat systemic diseases. However, the relatively low levels of gene expression obtained from using naturally occurring promoters, including the strong cytomegalovirus (CMV) enhancer/promoter (E/P), have limited the use of muscle as a target tissue. The relatively weak simian virus 40 (SV40) enhancer is known to have dual functions promoting localization of DNA to the nucleus and activating transcription. An SV40 enhancer incorporated either at the 5′ end of CMV E/P or the 3′ end of the polyadenylation site gave as much as a 20-fold increase in the level of exogenous gene expression in muscle in vivo, compared with expression observed with CMV E/P alone. The minimum requirement for this enhancement is a single copy of a 72-bp element of the SV40 enhancer, in combination with either the CMV E/P or skeletal actin (SkA) promoter. Enhancement of gene expression in muscle by this SV40 enhancer was also observed by using the powerful electroporation delivery. However, the SV40 enhancer does not increase the level of CMV E/P driven reporter gene expression in dividing tumor cells in vivo and in the dividing myoblast cell C2C12 in vitro. The data suggest that including this enhancer in the plasmid will enhance the level of gene production for muscle-based gene therapy.

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

Similar content being viewed by others

References

  1. Wolff JA et al. Direct gene transfer into mouse muscle in vivo Science 1990 247: 1465–1468

    Article  CAS  Google Scholar 

  2. Wolff JA et al. Long-term duration of plasmid DNA and foreign gene expression in mouse muscle Hum Mol Genet 1992 1: 363–369

    Article  CAS  Google Scholar 

  3. Danko I et al. Pharmacological enhancement of in vivo foreign gene expression in muscle Gene Therapy 1994 1: 114–121

    CAS  Google Scholar 

  4. Coney L et al. Facilitated DNA inoculation induces anti-HIV-1 immunity Vaccine 1994 12: 1545–1550

    Article  CAS  Google Scholar 

  5. Li X, Eastman EM, Schwartz RJ, Draghia-Akli R . Synthetic muscle promoters: activities exceeding naturally occurring regulatory sequences Nat Biotechnol 1999 17: 241–245

    Article  CAS  Google Scholar 

  6. Monahan PE et al. Direct intramuscular injection with recombinant AAV vectors results in sustained expression in a dog model of hemophilia Gene Therapy 1998 5: 40–49

    Article  CAS  Google Scholar 

  7. Sabelnikov AG . Nucleic acid transfer through cell membranes: towards the underlying mechanisms Prog Biophys Mol Biol 1994 162: 119–152

    Article  Google Scholar 

  8. Muramatsu T, Nakamura A, Park HM . In vivo electroporation: a powerful and convenient means of nonviral gene transfer to tissues of living animals Int J Mol Med 1998 1: 55–62

    CAS  Google Scholar 

  9. Neumann E, Kakorin S, Toensing K . Fundamentals of electroporative delivery of drugs and genes Bioelectrochem Bioenerg 1999 48: 3–16

    Article  CAS  Google Scholar 

  10. Dean DA . Import of plasmid DNA into the nucleus is sequence specific Exp Cell Res 1997 230: 293–302

    Article  CAS  Google Scholar 

  11. Vacik J, Dean BS, Zimmer WE, Dean DA . Cell-specific nuclear import of plasmid DNA Gene Therapy 1999 6: 1006–1014

    Article  CAS  Google Scholar 

  12. Dean DA, Dean BS, Muller S, Smith LC . Sequence requirements for DNA plasmid nuclear import Exp Cell Res 1999 253: 713–722

    Article  CAS  Google Scholar 

  13. Abruzzese RV et al. Ligand-dependent regulation of plasmid-based transgene expression in vivo Hum Gene Ther 1999 10: 1499–1507

    Article  CAS  Google Scholar 

  14. Rols MP et al. In vivo electrically mediated protein and gene transfer in murine melanoma Nat Biotechnol 1998 6: 168–171

    Article  Google Scholar 

  15. 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 

  16. Mathiesen I . Electropermeabilization of skeletal muscle enhances gene transfer in vivo Gene Therapy 1999 6: 508–514

    Article  CAS  Google Scholar 

  17. Kumar R, Firak TA, Schroll CT, Subramanian KN . Activation of gene expression is adversely affected at high multiplicities of linked simian virus 40 enhancer Proc Natl Acad Sci USA 1986 83: 3199–3203

    Article  CAS  Google Scholar 

  18. Graessmann M et al. Helper activity for gene expression, a novel function of the SV40 enhancer Nucleic Acids Res 1989 17: 6603–6612

    Article  CAS  Google Scholar 

  19. Doll RF et al. Comparison of promoter strengths on gene delivery into mammalian brain cells using AAV vectors Gene Therapy 1996 3: 437–447

    CAS  Google Scholar 

  20. Li S et al. Increased level and duration of expression in muscle by co-expression of a transactivator using plasmid systems Gene Therapy 1999 6: 2005–2011

    Article  CAS  Google Scholar 

  21. Mumper RJ et al. Protective interactive noncondensing (PINC) polymers for enhanced plasmid distribution and expression in rat skeletal muscle J Control Rel 1998 52: 191–203

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Otolaryngology-Head and Neck Surgery, University of Arkansas Medical School, Little Rock, AR, USA

    S Li

  2. Valentis, Inc., The Woodlands, TX, USA

    F C MacLaughlin, J G Fewell, M Gondo, J Wang, F Nicol & L C Smith

  3. Division of Pulmonary and Critical Care Medicine, Northwestern University, Chicago, IL, USA

    D A Dean

Authors
  1. S Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F C MacLaughlin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J G Fewell
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M Gondo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. F Nicol
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. D A Dean
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. L C Smith
    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

Li, S., MacLaughlin, F., Fewell, J. et al. Muscle-specific enhancement of gene expression by incorporation of SV40 enhancer in the expression plasmid. Gene Ther 8, 494–497 (2001). https://doi.org/10.1038/sj.gt.3301419

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301419

Keywords

This article is cited by

Search

Advanced search

Quick links