*2.3. Evaluation of Target Gene Transcription*

Specific mRNA synthesis of target genes was evaluated in eukaryotic cells 293T transfected with pEV.CTL and pEV.Th using MATra-A reagent according to the manufacturer's instruction (PromoKine, Heidelberg, Germany). Cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) with 10% FBS. 48 h after transfection mRNAs were isolated from cells with a kit for RNA isolation (Promega, Madison, WI, USA). Before reverse transcription all RNA samples were treated with RNase-free DNase. cDNAs were obtained by reverse transcription using RevertAid H Minus First Strand cDNA Synthesis Kit (Thermo Scientific, Berlin, Germany). Further, the obtained cDNA carried out PCR with the use of specific primers to gene *EV.CTL* (fCTL—AACTCAGGCACTCTTCCTGC, rCTL—TCGTACCGGAATCTCAGGGT) and gene *EV.Th* (fTh—ACGTTGACAAGCTGAGGAGG, rTh—GAGAGTCCTCAGCCCAGAGA). The amplification product was analyzed by electrophoresis in 1% agarose gel.
