{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T12:30:00Z","timestamp":1762605000679},"reference-count":49,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2003,8,11]],"date-time":"2003-08-11T00:00:00Z","timestamp":1060560000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Journal of Gene Medicine"],"published-print":{"date-parts":[[2003,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>In neonatal and adult rodent retina, substantial lentiviral vector expression has been detected primarily in retinal pigment epithelium (RPE), except in very young animals (2\u20135 days post\u2010natal). In non\u2010retinal tissues, studies of lentiviral vectors have utilized various controls. Among the most stringent are class I integrase mutants, which selectively block the integration reaction while leaving all other <jats:italic>gag<\/jats:italic>\/<jats:italic>pol<\/jats:italic>\u2010encoded functions intact. For HIV\u20101 vectors injected into brain, these have been used to simultaneously control for pseudotransduction and verify that long\u2010term expression requires integration. Such experiments compare particles that differ only in a single amino acid within a single enzyme that forms a very small molar fraction of the virion. Class I integrase mutants have not been described for feline immunodeficiency virus (FIV) integrase, or tested in the eye for any lentiviral vector.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We compared subretinally and intravitreally injected FIV vectors and followed animals for up to 7 months, a duration that exceeds prior studies. We also compared the wild\u2010type (WT) vector with one incorporating a single class I amino acid mutation in FIV integrase (D66V). A mock vector (packaging construct absent) was an alternative control. All vectors were vesicular stomatitis virus glycoprotein G (VSV\u2010G)\u2010pseudotyped and were injected on day 7 of life. One group of animals received either subretinal or intravitreal injections of WT vector in the right eyes. Control left eyes were injected with mock vector. These animals were sacrificed at 2 or 7 days post\u2010injection. A second group received subretinal injections of either WT vector or equivalent D66V vector (reverse transcriptase\u2010normalized to WT), and were analyzed after 2, 3 and 7 months. All eyes were scored for marker gene (\u03b2\u2010galactosidase) expression by an observer blinded to vector assignments.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Subretinal FIV vector injections were much more effective than intravitreal injections. The RPE was the principal retinal layer transduced by the WT vector, and at least 50% of the area of the retina expressed the marker gene at 3 and 7 months. Occasional cells in inner retinal layers also expressed \u03b2\u2010galactosidase at these time points. The sustained retinal expression produced by subretinally injected vector was blocked by the D66V mutation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>These results show that class I integrase mutant FIV vectors are useful control vectors, and that VSV\u2010G\u2010pseudotyped FIV vectors produce extensive retinal expression for at least 215 days, the longest duration yet reported for lentiviral vectors in retina. Transgene expression is mostly restricted to RPE after post\u2010natal day 7 in rats, suggesting that FIV vectors could be used to target RPE for gene therapy. Copyright \u00a9 2003 John Wiley &amp; Sons, Ltd.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jgm.447","type":"journal-article","created":{"date-parts":[[2003,12,3]],"date-time":"2003-12-03T08:00:38Z","timestamp":1070438438000},"page":"1009-1017","source":"Crossref","is-referenced-by-count":30,"title":["Comparison of wild\u2010type and class I integrase mutant\u2010FIV vectors in retina demonstrates sustained expression of integrated transgenes in retinal pigment epithelium"],"prefix":"10.1002","volume":"5","author":[{"given":"Nils","family":"Loewen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David 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