CLONING AND TRANSIENT EXPRESSION OF CYTOPROTECTIVE FACTOR, HO-1, IN MESENCHYMAL STEM CELLS USING THE ADENOVIRAL EXPRESSION SYSTEM THROUGH GATEWAY TECHNOLOGY

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HAMEDI ASL P.; HALABIAN R.; MOHAMADZADEH M.; MOHAMMADIPOUR M.; BAKHSHANDEH Z.; HAMEDI ASL D.; KIANI A.A.; JALILI M.; AMIRIZADEH N.; HABIBI ROUDKENAR M.

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Journal: The Scientific Journal of Iranian Blood Transfusion Year:2012 | Volume:9 | Issue:3 (36) Page(s): 214-225

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Title

CLONING AND TRANSIENT EXPRESSION OF CYTOPROTECTIVE FACTOR, HO-1, IN MESENCHYMAL STEM CELLS USING THE ADENOVIRAL EXPRESSION SYSTEM THROUGH GATEWAY TECHNOLOGY

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Keywords

MESENCHYMAL STEM CELLSQ3

HEME OXYGENASE-1Q3

GENETIC ENGINEERINGQ3

ADENOVIRUSESQ3

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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APA: Copy

HAMEDI ASL, P., HALABIAN, R., MOHAMADZADEH, M., MOHAMMADIPOUR, M., BAKHSHANDEH, Z., HAMEDI ASL, D., KIANI, A.A., JALILI, M., AMIRIZADEH, N., & HABIBI ROUDKENAR, M.. (2012). CLONING AND TRANSIENT EXPRESSION OF CYTOPROTECTIVE FACTOR, HO-1, IN MESENCHYMAL STEM CELLS USING THE ADENOVIRAL EXPRESSION SYSTEM THROUGH GATEWAY TECHNOLOGY. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), 9(3 (36)), 214-225. SID. https://sid.ir/paper/78323/en

Vancouver: Copy

HAMEDI ASL P., HALABIAN R., MOHAMADZADEH M., MOHAMMADIPOUR M., BAKHSHANDEH Z., HAMEDI ASL D., KIANI A.A., JALILI M., AMIRIZADEH N., HABIBI ROUDKENAR M.. CLONING AND TRANSIENT EXPRESSION OF CYTOPROTECTIVE FACTOR, HO-1, IN MESENCHYMAL STEM CELLS USING THE ADENOVIRAL EXPRESSION SYSTEM THROUGH GATEWAY TECHNOLOGY. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)[Internet]. 2012;9(3 (36)):214-225. Available from: https://sid.ir/paper/78323/en

IEEE: Copy

P. HAMEDI ASL, R. HALABIAN, M. MOHAMADZADEH, M. MOHAMMADIPOUR, Z. BAKHSHANDEH, D. HAMEDI ASL, A.A. KIANI, M. JALILI, N. AMIRIZADEH, and M. HABIBI ROUDKENAR, “CLONING AND TRANSIENT EXPRESSION OF CYTOPROTECTIVE FACTOR, HO-1, IN MESENCHYMAL STEM CELLS USING THE ADENOVIRAL EXPRESSION SYSTEM THROUGH GATEWAY TECHNOLOGY,” THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), vol. 9, no. 3 (36), pp. 214–225, 2012, [Online]. Available: https://sid.ir/paper/78323/en

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