Comparative evaluation of the transgene expression efficiency provided by the model genetic constructs of different structure

Abstract

Comparative evaluation of transgene expression efficiency provided by genetic constructs of different structures is an important stage in the development of new and optimization of existing expression vectors. However, there is no universal approach at present for correct comparison of expression efficiency of vectors of different structure. The goal of this work was to develop an experimental system for comparative evaluation of the expression efficiency provided by nonviral genetic vectors of various size and topology in human cell cultures. This system is based on the gene for the green fluorescence protein used as a reporter, flow cytometry methods for the evaluation of expression levels, and quantitative PCR for the adequate selection of transfection conditions. This system was tested with two model constructs: a linear DNA molecule and a circular plasmid.