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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References
Chen, Z.Y., He, C.Y., Ehrhardt, A. & Kay, M.A. Mol. Ther. 8, 495–500 (2003).
Huang, M. et al. Circulation 120, S230–S237 (2009).
Jia, F.J. et al. Nat. Methods 7, 197–199 (2010).
Riu, E., Chen, Z.Y., Xu, H., He, C.Y. & Kay, M.A. Mol. Ther. 15, 1348–1355 (2007).
Tan, Y., Li, S., Pitt, B.R. & Huang, L. Hum. Gene Ther. 10, 2153–2161 (1999).
Chen, Z.Y., He, C.Y. & Kay, M.A. Hum. Gene Ther. 16, 126–131 (2005).
Darquet, A.M., Cameron, B., Wils, P., Scherman, D. & Crouzet, J. Gene Ther. 4, 1341–1349 (1997).
Bigger, B.W. et al. J. Biol. Chem. 276, 23018–23027 (2001).
Mayrhofer, P., Blaesen, M., Schleef, M. & Jechlinger, W. J. Gene Med. 10, 1253–1269 (2008).
Khlebnikov, A., Datsenko, K.A., Skaug, T., Wanner, B.L. & Keasling, J.D. Microbiology 147, 3241–3247 (2001).
Morgan-Kiss, R.M., Wadler, C. & Cronan, J.E. Jr. Proc. Natl. Acad. Sci. USA 99, 7373–7377 (2002).
Yu, D. et al. Proc. Natl. Acad. Sci. USA 97, 5978–5983 (2000).
Thorpe, H.M. & Smith, M.C. Proc. Natl. Acad. Sci. USA 95, 5505–5510 (1998).
Minaeva, N.I. et al. BMC Biotechnol. 8, 63 (2008).
Sallam, K.I., Tamura, N., Imoto, N. & Tamura, T. Appl. Environ. Microbiol. 76, 2531–2539 (2010).
Datsenko, K.A. & Wanner, B.L. Proc. Natl. Acad. Sci. USA 97, 6640–6645 (2000).
Cherepanov, P.P. & Wackernagel, W. Gene 158, 9–14 (1995).
Stoll, S.M., Ginsburg, D.S. & Calos, M.P. J. Bacteriol. 184, 3657–3663 (2002).
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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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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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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DOI: https://doi.org/10.1038/nbt.1708
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