Accommodation of foreign genes into the Sendai virus genome: sizes of inserted genes and viral replication

Abstract

Sendai virus (SeV) is an enveloped virus with a negative sense genome RNA of about 15.3 kb. We previously established a system to recover an infectious virus entirely from SeV cDNA and illustrated the feasibility of using SeV as a novel expression vector. Here, we have attempted to insert a series of foreign genes into SeV of different lengths to learn how far SeV can accommodate extra genes and how the length of inserted genes affects viral replication in cells cultured in vitro and in the natural host, mice. We show that a gene up to 3.2 kb can be inserted and efficiently expressed and that the replication speed as well as the final virus titers in cell culture are proportionally reduced as the inserted gene length increases. In vivo, such a sizedependent effect was not very clear but a remarkably attenuated replication and pathogenicity were generally seen. Our data further confirmed reinforcement of foreign gene expression in vitro from the V(3 3) version of SeV in which the accessory V gene had been knocked out. Based on these results, we discuss the utility of SeV vector in terms of both efficiency and safety.z 1999 Federation of European Biochemical Societies.

SOC1 inhibits HPV-E7-mediated transformation by inducing degradation of E7 protein

“…Recombinant SeVs were constructed, as described (Kato et al, 1996;Sakai et al, 1999;Yonemitsu et al, 2000). In brief, 18 bp of spacer sequence 5 0 -(G)-CGGCCGCAGATCTTCACG-3 0 with a NotI restriction site (underlined) were inserted between the 5 0 nontranslated region and the initiation codon of the nucleoprotein (N) gene.…”

Skeletal muscle regeneration after insulin-like growth factor I gene transfer by recombinant Sendai virus vector

“…We investigated whether or not SeVvector-mediated hIGF-I gene transfer can induce morphological change to L6 cells, a neonatal rat myoblast cell line. The newly constructed recombinant SeV vector harboring human IGF-I (hIGF-I) gene, designated as hIGF-I/SeV, and the vector carrying ␤-galactosidase gene (LacZ/SeV) 16 were used and investigated to clarify the following: (1) the detection of hIGF-I expression in the cell culture supernatant (Western blotting analysis); (2) the kinetics of hIGF-I expression in the supernatant (ELISA assay); and (3) the morphological change in L6 cells. (1) In the L6 cells infected with hIGF-I/SeV with a multiplicity of infectivity (MOI) of 0.1, a polypeptide with an approximate molecular mass of 12-13 kDa reactive to the anti hIGF-I antibody was observed in the culture supernatant at 3 to 4 days after infection ( Figure 1a, lanes 3 and 4).…”

Recombinant Sendai virus provides a highly efficient gene transfer into human cord blood-derived hematopoietic stem cells

“…18,19 In brief, 18 bp of spacer sequence 5 0 -(G)-CGGCCGCAGATCTT-CACG-3 0 with a NotI restriction site were inserted between the 5' non-translated region and the initiation codon of the nucleoprotein (N) gene. This cloned SeV genome also contains a self-cleaving ribosome site from the antigenomic strand of the hepatitis delta virus.…”