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Françoise Haeseleer, Yoshikazu Imanishi, David A. Saperstein, Krzysztof Palczewski; Gene Transfer Mediated by Recombinant Baculovirus into Mouse Eye. Invest. Ophthalmol. Vis. Sci. 2001;42(13):3294-3300.
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© ARVO (1962-2015); The Authors (2016-present)
purpose. To determine the efficiency of baculoviruses (BVs) to transfer
recombinant genes in vivo into murine ocular tissues.
methods. Recombinant (r)BVs carrying fluorescent protein (FP) cDNA under the
control of cytomegalovirus (CMV) immediate early promoter were
constructed. Initially, cultured HEK293 and ARPE19 cells were infected
with these rBVs and analyzed for efficiency and stability of transgene
expression. The rBV-CMV green (G)FP was also injected into the
intravitreal and subretinal space of mouse eye. Mice were periodically
analyzed to determine the efficiency and stability of expression by
histologic examination under fluorescence microscopy. The effect of
rBV-CMV-GFP on the physiology of the retina was analyzed by
results. cDNAs encoding fluorescent proteins were efficiently transduced in
HEK293 and ARPE19 cells in vitro. GFP expression in vivo was observed
exclusively in retinal pigment epithelial (RPE) cells after subretinal
injections. Intravitreal injections of rBV resulted in GFP expression
in the corneal endothelium, lens, RPE, and retina. GFP expression was
observed for up to 14 days after injection. The infiltration of
macrophages, observed 2 days after injection in the area of GFP
transduction, had dissipated by day 8 after injection. No alteration in
ERG responses was observed 6 weeks after injection of rBV-CMV-GFP.
conclusions. BV efficiently transduces cultured RPE cells and many cell types in
vivo in the eye, including endothelial, epithelial, and neuronal cells.
BV may be a useful vector for transferring genes in cultured cells and
in vivo into ocular tissue.
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