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Last update: May 2021

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Human Nutrition Unit

Zone de texte éditable et éditée et rééditée

Electroporation

In vivo and in cellulo electroporation

Main characteristics of the NEPA21 electroporator (Sonidel) : New generation electroporator like the NEPA21 ( Figure 1 ) allows high efficiency transfection of hard to transfect cultured cells (muscle myotubes, primary cells, etc...) but also of most organs in vivo (brain, muscle, skin, liver, etc...). High efficiency transfection of plasmids High cellular viability No need for special buffers How does it work ? Transfection is achieved through electrical pulses with 2 main steps that can be adapted and optimized for each application. “Poring” is the first steps and opens the cellular membrane for allowing the penetration of nucleic acids. This step is achieved through high voltage (150-300 Volts In cultured cells) but during short periods (2-5 milliseconds). A succession of poring pulses is generally performed and different parameters can be modified, like a decay between the pulses and an inversion of polarity for increasing poring efficiency ( Figure 2 ). The “Transfer“ step allows the DNA to migrate into the cells thanks to a succession of mild voltage (30-60 Volts) but longer (50 milliseconds) electrical pulses. Again, parameters can be adapted to any cell type for achieving the best efficiency and viability. Probes for in vivo and in vitro electroporation While the principle is similar for any kind of electroporation, the probes used for achieving high efficiency electroporation are specific to any application. Cultured cells can be electroporated using a “tripod” electrode ( Figure 3 A ) while cells in suspension needs a cuvette. In living mice, electrodes are adapted to each organ ( Figure 3 B ). Pre-established programs are available, which can be used as a starting point for setting-up specific experiments. Contact : Daniel TAILLANDIER