Display of hepatitis B virus PreS1 peptide on bacteriophage T7 and its potential in gene delivery into HepG2 cells

• Main Authors: Tang, Kie Hie, Mohd Yusoff, Khatijah, Tan, Wen Siang
• Format: Article
• Language: English
• Published: Elsevier 2009
• Online Access: http://psasir.upm.edu.my/id/eprint/14097/1/Display%20of%20hepatitis%20B%20virus%20PreS1%20peptide%20on%20bacteriophage%20T7%20and%20its%20potential%20in%20gene%20delivery%20into%20HepG2%20cells.pdf; http://psasir.upm.edu.my/id/eprint/14097/

Hepatitis B is a major public health problem worldwide which may lead to chronic liver diseases, cirrhosis and hepatocellular carcinoma. An interaction between hepatitis B virus (HBV) envelope protein, particularly the PreS1 region, and a specific cell surface receptor is believed to be the initial step of HBV infection through attachment to hepatocytes. In order to develop a gene delivery system, bacteriophage T7 was modified genetically to display polypeptides of the PreS1 region. A recombinant T7 phage displaying amino acids 60-108 of the PreS1 region (PreS1(60-108)) was demonstrated to be most effective in transfecting HepG2 cells in a dose- and time-dependant manner. The phage genome was recovered from the cell lysate and confirmed by PCR whereas the infectious form of the internalized phage was measured by a plaque-forming assay. The internalized phage exhibited the appearance of green fluorescent dots when examined by immunofluorescence microscopy. Surface modification, particularly by displaying the PreS1(60-108) enhanced phage uptake, resulting in more efficient in vitro gene transfer. The ability of the recombinant phage to transfect HepG2 cells demonstrates the potential of the phage display system as a gene therapy for liver cancer.