A non-viral vector with transfection activity comparable to adenoviral transduction

ORCiD

Nejat Düzgüneş: 0000-0001-6159-1391

Department

Biomedical Sciences

Document Type

Conference Presentation

Conference Title

93rd General Session & Exhibition of the International Association for Dental Research (IADR)

Location

Boston, MA

Conference Dates

March 11-14, 2015

Date of Presentation

3-13-2015

Journal Title

Journal of Dental Research

Journal ISSN

0022-0345

Journal Volume Number

94

First Page

2009

Abstract

Objectives: To identify a highly efficient non-viral vector in comparison with adenoviral transduction for the delivery of therapeutic genes to oral cancer cells. While viral vectors are believed to be more efficient gene delivery vehicles, non-viral lipoplexes are advantageous because they are noninfectious, cost-effective, and easy to administer. A novel transfection reagent, TransfeXTM, shows extraordinary potential due to its high transfection efficiency, with low cytotoxicity. Methods: HeLa cervical carcinoma and HSC-3 human oral squamous cell carcinoma cells were seeded in 48-well culture plates one day before transduction or transfection, and used at ~85% confluence. Cells were transduced with Adenovirus CMV eGFP–RSV Luc at concentrations of 100, 1,000, or 10,000 viral particles per cell (vps/cell), or transfected with the cationic liposomal compound TransfeX at ratios of 2 and 4 µl TransfeX:1 µg DNA. Toxicity was measured with the Alamar Blue cell viability assay. Transfection efficacy was evaluated by measuring luciferase activity. Fluorescence microscopy and flow cytometry were used to examine the percentage of cells expressing GFP. Results: In HeLa cells, luciferase expression with TransfeX reached 52,140,000±2,200,818 RLU/ml compared to 14,442,000±629,708 RLU/ml after transduction with 10,000 vps/cell. In HSC-3 cells, transfection with TransfeX resulted in 559,467±30,624 RLU/ml, compared to 1,092,200±88,611 RLU/ml after transduction with 10,000 vps/cell. There was ~20% cytotoxicity at 4 µl TransfeX:1 µg DNA with both HeLa and HSC-3 cells. Viral transduction with HeLa cells caused no cytotoxicity, whereas HSC-3 cells showed ~20% cytotoxicity. Conclusions: TransfeX mediated 3.6-fold higher gene expression in HeLa cells than adenovirus; but was 2-fold less efficient in HSC-3 cells. TransfeX may be useful for gene therapy applications where adenovirus may be contraindicated. Therapeutic gene delivery to oral cancer cells may be achieved with the use of adenovirus. Future studies will examine the potential enhancement of adenoviral transduction by TransfeX.

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