Inhibition of Hepatitis B Virus Replication by Helper Dependent Adenoviral Vectors Expressing Artificial Anti-HBV Pri-miRs from a Liver-Specific Promoter
Figure 1
Schematic illustration of artificial pri-miR expression cassettes, recombinant HD Ads and HBV target sites. (a) Mono- or trimeric anti-HBV pri-miR sequences are transcribed under control of either a CMV or MTTR promoter. The cassettes also include an intron and polyA transcription termination signal. (b) The control HD Ad Δ28 vector contains no HBV-targeting cassettes but does include the CMV LacZ reporter cassette. Apart from ITRs and viral packaging signal (Ψ) the remainder of the vector DNA comprises stuffer sequence with all Ad open reading frames (ORFs) removed. (c) Anti-HBV HD Ads contain the mono- or trimeric artificial pri-miR expression cassettes under control of the CMV or MTTR Pol II promoters. (d) Tandem arrangement of a dimer of the hepatitis B virus genome, comprising 6.4 kb, showing sites targeted by antiviral artificial pri-miRs. The overlapping surface, core, polymerase, and HBx viral ORFs are indicated by labelled rectangles. The four arrows above the genome represent the HBV transcripts that are initiated from different viral promoter elements and a common 3′ end. Essential cis elements controlling transcription are indicated by arrowheads. The sites targeted by the artificial pri-miRs, indicated by the shaded rectangle, are located within the HBx ORF and are found in all of the viral transcript.
(a) HBV-targeting Pol II RNAi expression cassettes
