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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.

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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
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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.

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Correspondence to Mark A Kay.

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The authors declare no competing financial interests.

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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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