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

AUTHORS

Mohammad Mohammadzadeh 1 , Mehryar Habibi Roudkenar 2 , *

1 Dept. of Basic Sciences, School of Allied Medical Sciences, Gonabad University of Medical Sciences, Gonabad, Iran

2 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran

How to Cite: Mohammadzadeh M, Habibi Roudkenar M. Cloning and transient over-expression of the NRF2 gene in mesenchymal stem cells using adenoviral expression system based on the Gateway technology, J Kermanshah Univ Med Sci. 2013 ; 17(5):e77056.

ARTICLE INFORMATION

Journal of Kermanshah University of Medical Sciences: 17 (5); e77056
Published Online: August 29, 2013
Article Type: Research Article
Received: November 19, 2012
Accepted: April 30, 2013

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Abstract

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

Keywords

Mesenchymal Stem Cell NRF2 genetic engineering genetic vectors and cloning vector

© 2013, Journal of Kermanshah University of Medical Sciences. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

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