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URL: http://bloodjournal.ir/article-1-662-en.html
, R. Halabian , M. Mohammadzadeh , M. Mohammadipour , Z. Bakhshandeh , Dr. Hamedi Asl , A.A. Kianin , M.A. Jalili , N. Amirizadeh , M. Habibi Roudkenar *
Abstract
Background and Objectives
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.
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