Optimization of transfection of human coxsackie and adenovirus receptor into mammalian cells mediated by liposomes-base gene delivery

Baba, Sayang and Elengoe, Asita and Hussein, Abdul Rahim and Sugeng , Triwahyono and Draman, Samsul and Hamdan, Salehhuddin (2015) Optimization of transfection of human coxsackie and adenovirus receptor into mammalian cells mediated by liposomes-base gene delivery. Life Science Journal, 12 (2). pp. 152-158. ISSN 1097-8135

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Abstract

Cationic liposome-based reagents provide reliable and simple protocol for in vitro transfection in a broad range of mammalian cell types. High transfection efficacies and low cytotoxicity of a liposome-based transfection reagent was required to retain adequate viability and high level of transgene of transfected Chinese hamster ovary (CHO) cells for subsequent assay. Three commercial liposome-based transfection reagents were utilized and compared for their aptitude to transfect in-vitro CHO cells with reporter gene expressing the red fluorescent (pDsRed-N1) and fusion pDsRed-N1 plasmid containing human cDNA Coxsackie and Adenovirus Receptor (pCAR-DsRed). Xtreme HP Plasmid DNA transfection reagent (Roche) gave the highest transfection efficiencies compared to other tested reagents. The optimal Xtreme HP Plasmid DNA reagent had shown its higher transfection efficiency (39.25±1.00% MFI) and stable increment of DsRed-proportion cells with incubation period (24-72 hours), allowing sufficient yield and shortened the selection stably clone of successful transfected CHO-CARDsRed.

Item Type:	Article (Journal)
Additional Information:	5011/43121
Uncontrolled Keywords:	Coxsackie and Adenovirus Receptor, Chinese Hamster Ovary cell, Discoscoma red fluorescence protein, flow cytometry, inverted fluorescence microscope
Subjects:	R Medicine > R Medicine (General)
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Medicine > Department of Family Medicine (Effective: 1st January 2011)
Depositing User:	SR NUR FARHANA LATFI
Date Deposited:	26 May 2015 10:37
Last Modified:	26 May 2015 10:37
URI:	http://irep.iium.edu.my/id/eprint/43121

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