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Type of publication: research article
Type of publication (PDB): Straipsnis Clarivate Analytics Web of Science / Article in Clarivate Analytics Web of Science (S1)
Field of Science: Biofizika / Biophysics (N011)
Author(s): Šatkauskas, Saulius;Bureau, Michel F;Mahfoudi, Abderrahim;Mir, Lluis M
Title: Slow accumulation of plasmid in muscle cells : supporting evidence for a mechanism of DNA uptake by receptor-mediated endocytosis
Is part of: Molecular therapy. New York : Nature Publishing Group, 2001, Vol. 4, iss. 4
Extent: p. 317-323
Date: 2001
Keywords: Gene therapy;Electrogene therapy;Electrogenetherapy;Naked DNA;Plasmid DNA;Kinetics
Abstract: Intramuscular plasmid DNA injection results in long-term but low and variable expression of the injected genes. Optimization is difficult because the mechanism of naked DNA uptake by the cells in vivo is not yet determined. Here we used injections of plasmid DNA encoding luciferase to further characterize this mechanism. We analyzed the kinetics of naked DNA uptake by means of DNase I or heparin injections, using the level of luciferase expression as the indicator of DNA uptake. We demonstrated that in vivo heparin inhibits DNA uptake without affecting the expression of DNA internalized by means of electric pulses. Inhibition by heparin is dose dependent and compatible with the competition for the binding to a receptor. As shown also with DNase 1, DNA uptake by muscle cells is slow: a progressive accumulation of the DNA in the myofibers can be found for at least 4 hours after naked DNA injection. Physical presence of DNA molecules during the uptake period, but not later, was confirmed by the facilitation of DNA uptake with appropriate electric pulses. Therefore, uptake proceeds for the entire time during which intact DNA is present in the extracellular compartment. Our results support evidence for a DNA uptake mechanism based on receptor- mediated endocytosis
Affiliation(s): Gamtos mokslų fakultetas
Vytauto Didžiojo universitetas
Appears in Collections:1. Straipsniai / Articles
Universiteto mokslo publikacijos / University Research Publications

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