Electroporation method use an electrical field to create transient pores in cellular membrane which enables the intracellular delivery of charged molecules (for example – oligonucleotides, DNA, RNA, siRNA, proteins) to the cytoplasm (and nucleus) of the targeted cells. Delivery of genetic material in tissues using electric pulses for treating diseases is known as electroporation gene therapy. Discovered in 1982, electroporation is now widely used in cell and molecular biology laboratories for transtecting the genes of interest into cell lines to study the gene function, or inducing gene silencing by siRNA electroporation in vitro and in vivo. Electroporation services are offered by CRO laboratories.
Pre-optimized electroporation buffers and kits are available from Altogen Biosystems:
RNAi and siRNA Electroporation
RNA interference (RNAi) is experimental tool to induce gene silencing in cells (cytoplasm) via activating RISC complex to cleave target mRNA. Most examples using siRNAs have been associated to in vitro targets (cell lines). However, recently more studies focused on primary cells and in vivo animal models of human diseases. When electroporation is used to introduce siRNA or plasmid DNA into cells, high transfection efficiencies can be achieved when optimized electroporation conditions are utilized for specific cell line, primary cell type, or tissue type.
RNA interference process is activated by short interfering RNA (siRNA) that induce sequence-specific gene silencing and can be used to develop molecular therapeutics. siRNAs appear to be very promising new therapeutic agent, but it lacks safe and efficient delivery to the cell in vitro and tissues in vivo. siRNA electroporation technique can be applied to almost any tissue of a living animal, including tumors, skin, liver, kidney, artery, retina, cornea, or even brain.
Short interfering RNAs (siRNAs), a part of RNAi (RNA interference) present new gene silencing tool with therapeutic potential due to siRNA capacity to induce strong, sequence-specific gene silencing in cells and tissues. Electroporation of siRNA in vivo and in vitro represent highly efficient siRNA delivery method. Many studies demonstrated that gene silencing was efficiently obtained in vivo in an adult mouse with chemically synthesized siRNA after its electrical delivery in muscles. The associated gene silencing was measured in the same animal and lasted over 12 days.