Heme oxegenase1 (HO-1) is one of the potent cytoprotective factors. The goal of this study was to perform cloning and transient over expression of the human HO-1 gene in mesenchymal stem cells (MSCs) using the adenoviral expression system based on the gateway technology.
Materials and Methods
In order to induce expression of HO-1, A549 cell lines were exposed to UV for 1 hour. The full length cDNA of HO-1 was isolated and cloned into pENTR TOPO/D vector by TOPO cloning reaction. To construct the expression clone, a LR recombination reaction was carried out between the entry clone and destination vector, pAd/CMV/V5-DEST. The recombinant virus was produced in the appropriate mammalian cell line. MSCs were infected by the recombinant virus expressing HO-1.
Results
The results showed that human recombinant HO-1 was successfully cloned and the accuracy of the gene and its frame in the vector were confirmed by DNA sequencing. Expression of HO-1 in MSCs was confirmed by RT-PCR and western blot analysis. The results indicated that the expression of HO-1 is transient.
Conclusions
Transient expression of human HO-1 gene in MSCs by using adenovirus expression system may be considered as an efficient gene transfer strategy into MSCs in order to promote stem cell therapy.
Hamedi Asl P, Halabian R, Mohammadzadeh M, Mohammadipour M, Bakhshandeh Z, Hamedi Asl D, et al . Cloning and transient expression of cytoprotective factor, HO-1, in mesenchymal stem cells using the adenoviral expression system through Gateway technology. Sci J Iran Blood Transfus Organ 2012; 9 (3) :214-225 URL: http://bloodjournal.ir/article-1-662-en.html