Recombinant Sendai virus vectors deleted in both the matrix and the fusion genes: efficient gene transfer with preferable properties.
BACKGROUND
Sendai virus (SeV) is a new type of cytoplasmic RNA vector, which infects and replicates in most mammalian cells, directs high-level expression of the genes on its genome and is free from genotoxicity. In order to improve this vector, both the matrix (M) and fusion (F) genes were deleted from its genome.


METHODS
For the recovery of the M and F genes-deleted SeV (SeV/DeltaMDeltaF), the packaging cell line was established by using a Cre/loxP induction system. SeV/DeltaMDeltaF was characterized and compared with wild-type and F or M gene-deleted SeV vectors in terms of transduction ability, particle formation, transmissible property and cytotoxicity.


RESULTS
SeV/DeltaMDeltaF was propagated in high titers from the packaging cell line. When this vector was administered into the lateral ventricle and the respiratory tissue, many of the ependymal and epithelial cells were transduced, respectively, as in the case of wild-type SeV. F gene-deletion made the SeV vector non-transmissible, and M gene-deletion worked well to inhibit formation of the particles from infected cells. Simultaneous deletions of these two genes in the same genome resulted in combining both advantages. That is, both virus maturation into particles and transmissible property were almost completely abolished in cells infected with SeV/DeltaMDeltaF. Further, the cytopathic effect of SeV/DeltaMDeltaF was significantly attenuated rather than that of wild type in vitro and in vivo.


CONCLUSIONS
SeV/DeltaMDeltaF is an advanced type of cytoplasmic RNA vector, which retains efficient gene transfer, gains non-transmissible properties and loses particle formation with less cytopathic effect.
Inoue
Makoto
M
DNAVEC Research Inc., 1-25-11 Kannondai, Tsukuba-shi, Ibaraki 305-0856, Japan. inoue@dnavec.co.jp
Tokusumi
Yumiko
Y
Ban
Hiroshi
H
Shirakura
Masayuki
M
Kanaya
Takumi
T
Yoshizaki
Mariko
M
Hironaka
Takashi
T
Nagai
Yoshiyuki
Y
Iida
Akihiro
A
Hasegawa
Mamoru
M
D000818:Animals; D002412:Cations; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D005821:Genetic Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D005849:Gerbillinae; D049452:Green Fluorescent Proteins / Q000378:metabolism; D000882:Haplorhini; D007668:Kidney / Q000166:cytology; D007700:Kinetics; D008027:Light; D008081:Liposomes / Q000737:chemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016366:Open Reading Frames; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D012542:Scattering, Radiation; D029082:Sendai virus / Q000235:genetics*; D013997:Time Factors
D002412:Cations; D018076:DNA, Complementary; D008081:Liposomes; D049452:Green Fluorescent Proteins; D012313:RNA