Fig. 7
From: Multi-targeted gene silencing strategies inhibit replication of Canine morbillivirus
Schematic representation of multiple shRNA-expressing adenovirus construct. (a) The multiple shRNA-expressing cassette are transcribed under the control of U6 promoters. The cassette also includes polyT transcription termination signals (T) and spacer blocks (b) of 100 nt between shRNA-coding regions to prevent potential cross-interference. (b) Multi-shRNA expression cassette was cloned into the E1 region of an E1/E3-deleted Ad5 vector plasmid by restriction enzyme sites (Clal-Xbal). A recombinant adenovirus with scrambled shRNA sequence (Ad5SCR) was generated and used as a negative control. (c, d) Ad5-vector particles were produced by transfecting Pacl-linearized pAd5Ni(1–3) clone. HEK293 cells were transfected with pAd5Ni(1–3) and after 7–10 days, isolated plaques of recombinant Ad5 were picked to obtain the monoclonal virus, and the Ad5Ni(1–3) virus was further identified by PCR and sequencing. (d) Ad5Ni(1–3) transduction and cytopathic effects observed at days 7 and 10 (40X total magnification)