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:: Volume 21, Issue 4 (7-2016) ::
__Armaghane Danesh__ 2016, 21(4): 372-381 Back to browse issues page
Design Methods of Cell Adhesion Proteins Based on ELISA Usable in-vitro in Gene Therapy
E Hosseini1 , SY Hosseini2 , T Hashempour3 , MR Fattahi4 , M Sadeghizadeh 5
1- Department of Molecular Genetics, Tarbiat Modares University, Tehran, Iran,
2- Department of Bacteriology and Virology, Shiraz University of Medical Sciences, Shiraz, Iran
3- Department of Medical Virology, Alborzi Clinical Microbiology Research Center, Namazi Hospital, Shiraz University of Medical Sciences, Shiraz, Iran,
4- Gastroenterohepatology Research Center (GEHRC), Shiraz University of Medical Sciences, Shiraz, Iran
5- Department of Molecular Genetics, Tarbiat Modares University, Tehran, Iran, , sadeghma@modares.ac.ir
Abstract:   (4477 Views)

Background & aim: One of the strategies to improve the therapeutic gene is targeting gene therapy. A method which can be considered, is adding code sequences peptide or protein with high tendency to target cells and secreting the therapeutic gene encodes a protein. However, evaluating the effectiveness of such changes in the targeted cell binding protein gene product with the usual therapeutic methods produced in prokaryotic system is directly impossible. The purpose of this study was to evaluate the design methods of cell adhesion proteins based on ELISA usable in-vitro in gene therapy.

Methods: In order to target the therapeutic gene Mda-7 by using genetic engineering, peptide coding sequence RGD4C with the tendency to cancerous cell surface integrin were inserted shortly after the artificial signal peptide sequence and the N-terminal coding region of the protein. Then, the modified and unmodified cDNA eukaryotic expression vector pCDNA3.1 were matched. Vectors were transfected in HEK-293 cell line. Then Mda-7 secreted expression levels were measured in cell culture by ELISA. After adjusting the protein concentration of Mda-7 and RGD.Mda-7, in cells transfected media, they were used as a source of protein. Reduce the concentration of these genes was assessed two hours after exposure to the integrin cell lines with HepG2, M21 and lacking integrin Saos-2  were also determined by ELISA. The present study was conducted three times independently.  Data were analyzed using t-test.

Results: Statistical analysis of the results suggested that the gene product of the gene product RGD.Mda-7 and Mda-7 to connect to HepG2 cells and M21 were more likely to have integrin. While binding to the cell lines of Saos-2, no significant difference were observed.

Conclusions: It seems the present ELISA based method was a suitable strategy for cell attachment assay in gene therapy research.

Keywords: Cell adhesion assays, ELISA, RGD.Mda-7, RGD
Full-Text [PDF 354 kb]   (1738 Downloads)    
Type of Study: Research | Subject: General
Received: 2016/07/13 | Accepted: 2016/07/13 | Published: 2016/07/13
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Hosseini E, Hosseini S, Hashempour T, Fattahi M, Sadeghizadeh M. Design Methods of Cell Adhesion Proteins Based on ELISA Usable in-vitro in Gene Therapy. armaghanj 2016; 21 (4) :372-381
URL: http://armaghanj.yums.ac.ir/article-1-1416-en.html


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Volume 21, Issue 4 (7-2016) Back to browse issues page
ارمغان دانش Armaghane Danesh
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