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A robust system for production of minicircle DNA vectors

Nature Biotechnology volume 28, pages 1287–1289 (2010)Cite this article

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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.

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

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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

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
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  2. Cheng-Yi He
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  3. Zhi-Ying Chen
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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.

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The authors declare no competing financial interests.

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Supplementary Text and Figures

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

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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

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