Skip to main content
SpringerLink
Log in

A robust system for production of minicircle DNA vectors

  • Brief Communication
  • Published:

From Nature Biotechnology

View current issue Submit your manuscript

Abstract

Minicircle DNA vectors allow sustained transgene expression in quiescent cells and tissues. To improve minicircle production, we genetically modified Escherichia coli to construct a producer strain that stably expresses a set of inducible minicircle-assembly enzymes, ΦC31 integrase and I-SceI homing endonuclease. This bacterial strain produces purified minicircles in a time frame and quantity similar to those of routine plasmid DNA preparation, making it feasible to use minicircles in place of plasmids in mammalian transgene expression studies.

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

Figure 1: Comparison of the present and previous minicircle systems.
Figure 2: Improvement in minicircle quality and quantity.

Similar content being viewed by others

References

  1. Chen, Z.Y., He, C.Y., Ehrhardt, A. & Kay, M.A. Mol. Ther. 8, 495–500 (2003).

    Article  CAS  Google Scholar 

  2. Huang, M. et al. Circulation 120, S230–S237 (2009).

    Article  CAS  Google Scholar 

  3. Jia, F.J. et al. Nat. Methods 7, 197–199 (2010).

    Article  CAS  Google Scholar 

  4. Riu, E., Chen, Z.Y., Xu, H., He, C.Y. & Kay, M.A. Mol. Ther. 15, 1348–1355 (2007).

    Article  CAS  Google Scholar 

  5. Tan, Y., Li, S., Pitt, B.R. & Huang, L. Hum. Gene Ther. 10, 2153–2161 (1999).

    Article  CAS  Google Scholar 

  6. Chen, Z.Y., He, C.Y. & Kay, M.A. Hum. Gene Ther. 16, 126–131 (2005).

    Article  CAS  Google Scholar 

  7. Darquet, A.M., Cameron, B., Wils, P., Scherman, D. & Crouzet, J. Gene Ther. 4, 1341–1349 (1997).

    Article  CAS  Google Scholar 

  8. Bigger, B.W. et al. J. Biol. Chem. 276, 23018–23027 (2001).

    Article  CAS  Google Scholar 

  9. Mayrhofer, P., Blaesen, M., Schleef, M. & Jechlinger, W. J. Gene Med. 10, 1253–1269 (2008).

    Article  CAS  Google Scholar 

  10. Khlebnikov, A., Datsenko, K.A., Skaug, T., Wanner, B.L. & Keasling, J.D. Microbiology 147, 3241–3247 (2001).

    Article  CAS  Google Scholar 

  11. Morgan-Kiss, R.M., Wadler, C. & Cronan, J.E. Jr. Proc. Natl. Acad. Sci. USA 99, 7373–7377 (2002).

    Article  CAS  Google Scholar 

  12. Yu, D. et al. Proc. Natl. Acad. Sci. USA 97, 5978–5983 (2000).

    Article  CAS  Google Scholar 

  13. Thorpe, H.M. & Smith, M.C. Proc. Natl. Acad. Sci. USA 95, 5505–5510 (1998).

    Article  CAS  Google Scholar 

  14. Minaeva, N.I. et al. BMC Biotechnol. 8, 63 (2008).

    Article  Google Scholar 

  15. Sallam, K.I., Tamura, N., Imoto, N. & Tamura, T. Appl. Environ. Microbiol. 76, 2531–2539 (2010).

    Article  CAS  Google Scholar 

  16. Datsenko, K.A. & Wanner, B.L. Proc. Natl. Acad. Sci. USA 97, 6640–6645 (2000).

    Article  CAS  Google Scholar 

  17. Cherepanov, P.P. & Wackernagel, W. Gene 158, 9–14 (1995).

    Article  CAS  Google Scholar 

  18. Stoll, S.M., Ginsburg, D.S. & Calos, M.P. J. Bacteriol. 184, 3657–3663 (2002).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank P. Valdmanis for critical review of the manuscript. This work was supported by the US National Institutes of Health - HL064274 (M.A.K.). Bacterial strain BW27783 was a gift of Jay D. Keasling of the University of California at Berkeley. Plasmid placY A177C was obtained from John E. Cronan at the University of Illinois.

Author information

Author notes

  1. zychen98@stanford.edu

Authors and Affiliations

  1. Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California, USA

    Mark A Kay, Cheng-Yi He & Zhi-Ying Chen

  2. The Center for Clinical Science Research, Stanford University School of Medicine, Stanford, California, USA

    Mark A Kay, Cheng-Yi He & Zhi-Ying Chen

Authors
  1. Mark A Kay
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cheng-Yi He
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhi-Ying Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Z.-Y.C. planned and carried out the experiments. C.-Y.H. conducted a number of the experiments. M.A.K. and Z.-Y.C. discussed and planned the experimental strategies. M.A.K. and Z.-Y.C. wrote the manuscript.

Corresponding author

Correspondence to Mark A Kay.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Protocol, Supplementary Glossary and Supplementary Figs. 1–7 (PDF 4806 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kay, M., He, CY. & Chen, ZY. A robust system for production of minicircle DNA vectors. Nat Biotechnol 28, 1287–1289 (2010). https://doi.org/10.1038/nbt.1708

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt.1708

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation