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[Approach to transforming hepatitis B virus as a gene therapeutic vector]
Ju-qiang Han1, Da-rong Hu, Xue-ling Hu
1Institute of Hepatology, Beijing Military General Hospital, Beijing 100700, China.
Objective:
To evaluate the possibility of hepatitis B virus (HBV) as a vector in liver-targeting gene therapy.
Methods:
A fragment containing the small envelope gene of HBV was replaced with the reporter gene green fluorescent protein (GFP) to construct the recombinant HBV vector, which was transfected into HepG2 cells with liposome. The expression of GFP was observed with fluorescence microscope. The HBV cccDNA was testified using semi-nest PCR. The viral particles of the recombinant HBV in culture medium were detected by PCR as well as Southern blot.
Results:
The HBV vector carrying the interesting gene of GFP could express the functional protein in the transfected hepatocytes. However, the recombinant HBV vector was replication-deficient, which could not be packed and replicated in the hepatocytes to secrete mature recombinant HBV particles carrying the interesting gene of GFP when transfected solely but could when cotransfected with the recombinant and helper construct which lacked part of 5'-proximal HBV RNA packaging signal epsilon.
Conclusion:
It is possible that HBV is reconstructed as a liver-targeting vector for gene therapy.