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In vitro replication of hepatitis D virus using a new construct containing a cDNA dimer of HDV genome
Farida Behzadian1, Farzaneh Sabahi, Majid Sadeghizadeh
1Department of Virology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Background:
There is no cell line susceptible to hepatitis D virus (HDV) infection and capable of stable replication of its genome. Different genetic-based approaches have been introduced to initiate HDV replication events so far.
Methods:
In order to construct a replicative model for HDV made from a unique genome sequence, two monomeric units of HDV full-length cDNA were joined together through a four-step cloning scheme. The resulting vector (pcDNA3.1-D2) containing two tandem repeats of HDV cDNA under CMV promoter control was then used in transfection experiments into COS7 and HuH7 cell lines.
Results:
HDV replication markers including expression of hepatitis delta antigen (HDAg), the only HDV-specific antigen, and synthesis of antigenomic RNA were shown in both transfected cell lines, indicating initiation of HDV genome replication.
Conclusions:
Our results suggested that pcDNA3.1-D2, a vector containing a cDNA dimer of the HDV genome, originated from a unique full-length HDV molecule that is capable of replicating in cultured cells. This vector can be used conveniently for transfection experiments to study HDV molecular biology.
