Abstract
Minicircle DNA vectors are plasmid derivatives free of bacterial elements. These vectors are mostly provided from common plasmids via recombination by site-specific recombinases in E. coli. Absence of bacterial backbone in minicircle vectors results in high-level and persistent expression of transgene in comparison with conventional plasmids and provides promising vehicles for gene therapy and vaccination. Here we describe the production of replicative minicircle DNA vectors using the PBAD/araC system expressing ΦC31 integrase in E. coli.
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References
Chen ZY, Riu E, He C-Y et al (2008) Silencing of episomal transgene expression in liver by plasmid bacterial backbone DNA is independent of CpG methylation. Mol Ther 16:548–556
Kay MA (2011) State-of-the-art gene-based therapies: the road ahead. Nat Rev Genet 12:316–328
Gill D, Pringle I, Hyde S (2009) Progress and prospects: the design and production of plasmid vectors. Gene Ther 16:165–171
Lu J, Zhang F, Xu S et al (2012) The extragenic spacer length between the 5′ and 3′ ends of the transgene expression cassette affects transgene silencing from plasmid-based vectors. Mol Ther 20(11):2111–2119. doi:10.1038/mt.2012.65
Klinman DM (2004) Immunotherapeutic uses of CpG oligodeoxynucleotides. Nat Rev Immunol 4:249–259
Dong Y, Aied A, Li J et al (2013) An in vitro approach for production of non-scar minicircle DNA vectors. J Biotechnol 166:84–87
Sanei Ata-Abadi N, Dormiani K, Khazaie Y et al (2015) Construction of a new minicircle DNA carrying an enhanced green florescent protein reporter gene for efficient expression into mammalian cell lines. Mol Biol Rep 42:1175–1185
Kay MA, He CY, Chen ZY et al (2010) A robust system for production of minicircle DNA vectors. Nat Biotechnol 28:1287–1289
Darquet A, Cameron B, Wils P et al (1997) A new DNA vehicle for nonviral gene delivery: supercoiled minicircle. Gene Ther 4:1341–1349
Bigger BW, Tolmachov O, Collombet JM et al (2001) An araC-controlled bacterial Cre expression system to produce DNA minicircle vectors for nuclear and mitochondrial gene therapy. J Biol Chem 276:23018–23027
Nehlsen K, Broll S, Bode J et al (2006) Replicating minicircles: generation of nonviral episomes for the efficient modification of dividing cells. Gene Ther Mol Biol 10:233–244
Chen ZY, He CY, Ehrhardt A, Kay MA et al (2003) Minicircle DNA vectors devoid of bacterial DNA result in persistent and high-level transgene expression in vivo. Mol Ther 8:495–500
Jechlinger W, Azimpour Tabrizi C, Lubitz W et al (2004) Minicircle DNA immobilized in bacterial ghosts: in vivo production of safe non-viral DNA delivery vehicles. J Mol Microb Biotech 8:222–231
Chen ZY, He CY, Kay MA (2005) Improved production and purification of minicircle DNA vector free of plasmid bacterial sequences and capable of persistent transgene expression in vivo. Hum Gene Ther 16:126–131
Kobelt D, Schleef M, Schmeer M et al (2013) Performance of high quality minicircle DNA for in vitro and in vivo gene transfer. Mol Biotechnol 53:80–89
Forde GM, Ghose S, Slater NK et al (2006) LacO-lacI interaction in affinity adsorption of plasmid DNA. Biotechnol Bioeng 95:67–75
Ghose S, Forde GM et al (2004) Affinity adsorption of plasmid DNA. Biotechnol Prog 20:841–850
Hou XH, Guo XY, Chen Y et al (2015) Increasing the minicircle DNA purity using an enhanced triplex DNA technology to eliminate DNA contaminants. Mol Ther Methods Clin Dev 1:14062. doi:10.1038/mtm.2014.62
Argyros O, Wong SP, Fedonidis C et al (2011) Development of S/MAR minicircles for enhanced and persistent transgene expression in the mouse liver. J Mol Med 89:515–529
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Ata-abadi, N.S., Rezaei, N., Dormiani, K., Nasr-Esfahani, M.H. (2017). Production of Minicircle DNA Vectors Using Site-Specific Recombinases. In: Eroshenko, N. (eds) Site-Specific Recombinases. Methods in Molecular Biology, vol 1642. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7169-5_20
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DOI: https://doi.org/10.1007/978-1-4939-7169-5_20
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7169-5
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