Volume 20, Number 101 (11-2012)                   daneshvarmed 2012, 20(101): 13-26 | Back to browse issues page


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Rahbarizadeh F, Asle Rasouli N, Jafari Iri Sofla F, Ahmadvand D. Delivery of gene therapy construct by polyplexes nanoparticle (targeted polyethyleneimine) to colon cancer cells. daneshvarmed. 2012; 20 (101) :13-26
URL: http://daneshvarmed.shahed.ac.ir/article-1-605-en.html

Associate Professor Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University
Abstract:   (8095 Views)
 

Background and Objectives: The aim of this study was preparation of polyethyleneimine (PEI) nanoparticles targeted with anti-TAG72 nanobody for delivery of t-Bid gene construct into the human colonic adenocarcinoma cells.

 

Materials and Methods: First of all, purified recombinant nanobody was verified by SDS-PAGE, western blotting and ELISA. Targeted polyplexes were constructed by reacting polyethylene glycol (PEG) with PEI and covalent bonding of nanobody to nanoparticles. Surface charge and size of the resulting nanoparticles were evaluated by Malvern zeta sizer and nanosight instruments. Efficiency of constructed gene vector for killer gene delivery into the colonic adenocarcinoma cells was assessed using real time PCR assay.

 

Results: Production of nanoparticles with the average size of 168 nm and +7.18 surface charge was confirmed by nanosight and Malvern zeta sizer. Gel retardation assay verified the efficiency of carrier for pDNA complexation. Real time PCR results confirmed the targeted gene expression in the cancerous cell lines.

 

Conclusion: Results of this study confirmed the efficiency of targeted polyplexes in gene delivery and their potency for targeted gene delivery into the cells.

 
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Type of Study: Research |

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