| Autor: |
Hamedi Asl, P., Halabian, R., Mohamadzadeh, M., Mohammadipour, M., Bakhshandeh, Z., Hamedi Asl, D., Kiani, A. A., Jalili, M., Amirizadeh, N., Habibi Roudkenar, M. |
| Předmět: |
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| Zdroj: |
Scientific Journal of Iranian Blood Transfusion Organization; Autumn2012, Vol. 9 Issue 3, p214-225, 12p |
| Abstrakt: |
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. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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