Nucleic Acids Research, Vol 24, Issue 14 2697-2700, Copyright © 1996 by Oxford University Press
N Ramesh, ST Kim, MQ Wei, M Khalighi and WR Osborne
Bicistronic retroviral vectors were constructed containing the foot-and-
mouth disease virus (FMDV) internal ribosome entry site (IRES) followed by
the coding region of beta-galactosidase (beta-gal) or therapeutic genes,
with the selectable neomycin phosphotransferase gene under the control of
the viral long terminal repeat (LTR) promoter. LNFX, a vector with a
multiple cloning site 3' to foot-and-mouth disease virus IRES, was used to
construct vectors encoding rat erythropoietin (EP), rat granulocyte
colony-stimulating factor (G-CSF), human adenosine deaminase (ADA) and
beta-gal. In transduced primary rat vascular smooth muscle cells the
cytokines were expressed at high levels, similar to those obtained from
vectors employing the viral LTR promoter. LNFZ, a vector encoding beta-gal,
had a 10-fold increase in titer over that of LNPoZ, a comparable vector
containing the poliovirus (Po) internal ribosome entry site. Primary canine
vascular smooth muscle cells infected with LNFZ and LNPoZ expressed similar
activities of beta-gal and neomycin phosphotransferase (NPT). Overall,
these vectors had titers between 10(6) and 2 x 10(7) c.f.u./ml, indicating
that foot-and- mouth disease virus IRES provides high-titer bicistronic
vectors with high-level two gene expression.
ARTICLES
High-titer bicistronic retroviral vectors employing foot-and-mouth disease virus internal ribosome entry site
Department of Pediatrics, University of Washington School of Medicine Seattle 98195, USA.
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