Evaluation of HSV-1 Effects on Brain Cells Ion Channel Proteins Regulation in HEK293T cells

The HSV-1 can use its miRNAs, such as human miRs, to control the expression of ion channels, such as SCN8A, SLC22A2, SLC25A22, and KCNMA1. In this paper, the Target Scan software was used for targeting the ion channel genes by each of the LAT miRs and then two miRs were selected for the continuation of laboratory work. After the development of gene constructs containing each of the miRs and transfecting them into the HEK293T cell line, increased expression of miRs was confirmed by the Real-Time PCR technique, and increased expression of each miR developed by the LAT virus with plasmid transfect containing LAT and H4-5P, H5-5, H7, and H8 miRNAs was evaluated. In the next stage, the target genes were examined. To this end, first, the expression of the genes was evaluated separately using H2 and H3 miRs and based on Real-time PCR reaction. In order to check the effect of LAT transcript on any of the genes, the real-time reaction was performed on cDNAs made by transfection of LAT into HEK293T cells and the Luciferase test was performed on one of the genes (SLC22A2). The results indicate decreased SLC22A2 and SCN8A gene expression as tested by miR-H2 and LAT. As for the two other genes, an increase was observed in their expression, requiring the examination of other upstream and downstream genes. Luciferase assay showed that SLC22A2 is suppressed only by miR-H3, but for confirmation of this finding, it is necessary to check the effect of LAT transcript on SLC22A2. The populations of the study in these channels constitute part of the genes involved in neurological diseases in general and epilepsy in particular. Ionic channels can contribute to epilepsy through mutation or malfunctions, with SCN8A sodium channel playing a major role in such disorders.