In vitro studies on drug and gene transfer into cells

Abstract

Currently, the use of viral and chemical vectors is the most widespread in clinical gene therapy trials, but the side effects of these vectors, such as immunogenicity, toxicity, non-specificity or low efficacy, remain an unresolved problem. Compared to viral or chemical vector methods, electroporation of foreign genes is significantly simpler, cheaper and safer, but due to the lack of efficacy of tissue transfection, electroporation therapy has not yet reached clinical trials. Therefore, the delivery of foreign genes into cells or tissues by electropermeabilisation is a relatively new and extensively studied problem in medicine and biotechnology, and recent research in this field has focused on maximising the efficiency of gene electroporation [1,2]. The aim of study was theoretical analysis of gene transfer, evaluation of the properties and applicability of viral and non-viral vector types, and analysis of the electroporation methodology and its application in genetic engineering practice. Different electroporation parameters used to measure the viability of Chinese hamster ovary cells (CHO-K1) had different effects under different conditions (current strength, voltage and pulse duration), while maintaining the same energy content. The most effective cell viability in both plasmid and cell-free environments was found at 1800 V/cm; 61 μs. while stronger and shorter impulses had a greater negative impact on cell viability. Transfection during the electroporation process is relatively dependent on the parameters of the electrical pulse used - the most efficient gene transfer was recorded at 1400 V/cm; 100 μs, while delivery did not occur at other parameters. The total cell fluorescence data are regular to the transfection data, with a peak value obtained at 1400 V/cm and a pulse duration of 100 μs.