Cloning and transient over-expression of the NRF2 gene in mesenchymal stem cells using adenoviral expression system based on the Gateway technology

Increased mesenchymal stem cells (MSCs) survival after transplantation can improve their therapeutic potential. Therefore، manipulation of the MSCs with cytoprotective genes such as NF-E2 Related Factor-2 (NRF2) is important to improve cell therapy efficiency. The aim of this study was cloning and transient over-expression of the NRF2 gene in MSCs by adenoviral expression system. NRF2 was isolated from MSCs and cloned into pENTR/TOPO/D vector by TOPO cloning reaction. NRF2 was translocated from pENTR/TOPO/D vector to adenoviral vector using the Gateway technology. The recombinant adenovirus vector was transformed into 293A cells to package recombinant adenovirus particles. Recombinant adenoviruses harboring NRF2 infected MSCs. NRF2 was successfully isolated، cloned and its accuracy was confirmed by Deoxyribonucleic acid (DNA) sequencing. NRF2 over-expression was evaluated. This over-expression by adenoviral expression system was transient. NRF2 over-expression in MSCs with adenoviral expression system could be a valuable device because of its transient expression. Stable over-expression of genes in MSCs could not be permitted.