Reversible cell electroporation is a technique where an electric field is applied to cells to temporarily increase the permeability of their membranes, allowing the introduction of foreign molecules such as DNA, RNA, or proteins into the cells. The term “reversible” refers to the fact that the changes to the cell membrane are transient, and the membrane reseals itself after the electric field is removed, preserving cell viability and function.
The key to achieving reversible electroporation is to optimize the electric field parameters, such as voltage, pulse duration, and the number of pulses. These parameters depend on the cell type, size, and the molecules being introduced. When optimized correctly, reversible electroporation can efficiently deliver molecules into cells while maintaining high cell viability.
The general steps for reversible cell electroporation are as follows:
- Sample preparation: The target cells are mixed with the molecules to be introduced, such as plasmid DNA, siRNA, or proteins.
- Electric pulse application: The cell-molecule mixture is exposed to one or more electric pulses using an electroporation device. The electric field parameters must be optimized for the specific cell type and molecule to ensure efficient delivery and high cell viability.
- Recovery: Following the electric pulse application, the cells are allowed to recover in a suitable culture medium. During this time, the cell membrane reseals, and the introduced molecules can start to exert their effects within the cell, such as expressing a desired gene or silencing a target gene.
Reversible cell electroporation is widely used in various biological research and therapeutic applications, including gene transfer, gene editing, drug delivery, and cancer therapy. The technique can be applied in vitro (in cell culture), ex vivo (on isolated tissues or cells), or in vivo (directly within an organism). Proper optimization and execution of reversible electroporation are essential to minimize cell damage and maintain cell viability and function.