In vivo Gene electrotransfer: optimisation and mechanisms.
Identifieur interne : 000421 ( Main/Exploration ); précédent : 000420; suivant : 000422In vivo Gene electrotransfer: optimisation and mechanisms.
Auteurs : Franck AndréSource :
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Abstract
DNA electrotransfer is a promising solution to the problems induced by the use of viral transfer techniques, both in terms of therapeutic applications and as a tool for research. However, improvements have to be made concerning the transfection efficacy in the different cell lines, the knowledge of the underlying mechanisms and also the knowledge of the safety of the electrotransfer. In a preliminary study we have proved that transfection efficacy for DNA injection alone increased with the speed of injection, mainly due to receptor mediated endocytosis, but also partly to membrane permeabilisation. Afterward we have confirmed in vivo the permeabilising role of the short high voltage pulses (HVs) and the electrophoretic role of the long low voltage pulses (LVs), during the elctrotransfer. We have also shown the influence of a delay between the HV and the LV, on the efficacy and the toxicity of the pulses applied. Our study of the HV also tends to indicate that the tissue doesn't have to be entirely permeabilised to obtain a good electrotransfer. We have also confirmed in vivo that the smaller the cells are, the higher the voltage applied for the HV has to be. This is in agreement with Schwan's formula:
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