Hydrodynamics-based transfer of human apolipoprotein A-I gene to mice: Study of factors affecting the efficiency and duration of gene expression in the mouse liver

B. N. Akifiev, E. B. Dizhe, A. M. Efremov, D. A. Mogilenko, G. N. Oleinikova, I. A. Lapikov, O. Yu Zhdanova, O. V. Kidgotko, S. V. Orlov, A. P. Perevozchikov

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2 Scopus citations

Abstract

Human apolipoprotein A-I gene (apoA-I) inserted into a plasmid expression vector was transferred in vivo into C57Bl/6 mice using hydrodynamic injections into the tail vein. Two types of plasmid expression vectors were used: (1) pCMVcapoAI which contained cDNA of apoA-I driven by the human cytomegalovirus (CMV) early gene promoter and (2) pAlg, which contained a genomic locus of intron-containing apoA-I driven by its own extended 5′-regulatory region (APOAI). Hydrodynamic intravenous injections of both expression vectors led to the appearance of human apoA-I mRNA in the liver and human ApoA-I protein in the serum of injected mice. The dynamics of human ApoA-I content in the sera of mice injected with pCMVcapoAI and pAlg were different. When pCMVcapoAI was used, the concentration of human ApoA-I in mouse serum was maximal one day after injection and decreased to zero within the next two weeks. In the case of pAlg, the content of human ApoA-I in serum was maximal (up to 20 μg/ml) on days 5-7 after injection and then gradually decreased for several months (six months after injection, for example, it decreased to 25% of the maximal value). Experiments on "saved" pAlg plasmid isolated from the nuclei of hepatocytes 50 days after injection showed that the plasmid was retained for a long time in the form of an episome. A significant content of human ApoA-I in serum and its long-term persistence after injecting mice with pAlg may be accounted for by the properties of APOAI and/or the exon-intron structure of the apoA-I gene. Injecting mice with different variants of APOAI coupled with the luciferase gene did not lead to long-term expression of luciferase in the liver. It is concluded that the presence of introns in the apoA-I gene is required for its efficient and long-term expression after transfer to mice by means of hydrodynamic injections.

Original languageEnglish
Pages (from-to)921-928
Number of pages8
JournalMolecular Biology
Volume38
Issue number6
DOIs
StatePublished - Nov 2004

Keywords

  • C57Bl/6 mice
  • gene therapy
  • intravenous hydrodynamic injections
  • plasmid human apolipoprotein A-I gene expression vectors
  • significance of the exon-intron gene structure for long-term expression in vivo

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