Non-transmissible Sendai virus vector encoding c-myc suppressor FBP-interacting repressor for cancer therapy

Figure 2 Structures and procedures for generating fusion gene-deficient Sendai virus/dF/far up stream element-binding protein-interacting repressor (SeV/dF/FIR) or Sendai virus/dF/green fluorescent protein vectors (SeV/dF/GFP) from Sendai virus genome RNA. A: Schematic genome structures of wild-type (SeV) and fusion gene-deficient (SeV/dF; non-transmissible) vector carrying the human FIR (hFIR) gene or jellyfish green fluorescent protein (GFP). The open reading frame or the FIR or GFP gene was inserted with the SeV-specific transcriptional regulatory signal sequences. T7; T7 promoter, Rbz; hepatitis delta virus ribozyme sequence; B: Schematic representation of the procedure for generating the fusion gene-deficient SeV/dF/FIR or SeV/dF/GFP. SeV/dF/FIR or SeV/dF/GFP virus particles were propagated using fusion protein-expressing packaging cells (LLC-MK2/F7/A) after preparation in LLC-MK2 cells using the four plasmids driven by a recombinant vaccinia virus expressing T7 RNA polymerase which had been inactivated with psoralen and long-wave UV light (UV-vTF7-3); C: SeV/dF/FIR virus vectors were infected into HeLa cells and whole cell proteins were extracted for western blot analysis. SeV/dF/FIR expresses FIR proteins. FIR: FBP Interacting Repressor; FBP: FUSE-Binding protein; FUSE: Far Upstream Element; SeV: Sendai virus; NP: Nucleoprotein; P: Phosphoprotein; L: The catalytic subunit of the polymerase large protein forms a ribonucleoprotein complex (RNP) vector and was transfected separately with the SeV RNA. See Materials and Methods.