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Correlation between transgen expression and plasmid DNA loss in mouse liver
Author(s) -
Togashi Ryohei,
Harashima Hideyoshi,
Kamiya Hiroyuki
Publication year - 2013
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.2716
Subject(s) - plasmid , transgene , microbiology and biotechnology , biology , plasmid preparation , dna , gene , genetics , pbr322
Background Transgene expression from plasmid DNA is dependent on the expression efficiency per plasmid and the amount of intranuclear plasmid. In the present study, intranuclear dispositions of two types of plasmid DNAs (i.e. the pCpGfree and pLIVE plasmids) that maintain transgene expression in mouse liver were analyzed. In addition, the relationship between transgene expression and plasmid stability in the nucleus was examined. Methods First, the pCpGfree and pLIVE plasmid DNAs, bearing the mouse secreted alkaline phosphatase ( Seap ) gene, were administered into mouse liver by the hydrodynamics‐based method. Next, various Seap‐plasmid DNAs containing different promoters, upstream and downstream sequences, and backbones were injected into mice, and both SEAP expression and plasmid DNA amounts were monitored for 28 days. Results At the 14‐ and 28‐day time points, the amount of the pCpGfree plasmid DNA was one order of magnitude less than that of the pLIVE plasmid. Meanwhile, the expression efficiency per plasmid was one order of magnitude more efficient for the pCpGfree plasmid DNA. Moreover, the administration of various Seap‐plasmid DNAs revealed that negative correlations exist between plasmid stability and SEAP expression level. Conclusions The results obtained suggest that the pCpGfree plasmid is unstable from the viewpoint of quantity and maintains transgene expression by its high expression efficiency and also that transgene expression negatively affects the stability of plasmid DNA. Copyright © 2013 John Wiley & Sons, Ltd.