{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T01:06:25Z","timestamp":1773277585671,"version":"3.50.1"},"reference-count":64,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2005,1,17]],"date-time":"2005-01-17T00:00:00Z","timestamp":1105920000000},"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":[[2005,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>The goal of this study was to design improved regulatable lentivirus vector systems. The aim was to design tetracycline (tet)\u2010regulatable lentivirus vectors based on the Tet\u2010on system displaying low background expression in the absence of the doxycycline (DOX) inducer and high transgene expression levels in the presence of DOX.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We constructed a binary lentivirus vector system that is composed of a self\u2010inactivating (SIN) lentivirus vector bearing inducible first\u2010 or second\u2010generation tet\u2010responsive promoter elements (TREs) driving expression of a transgene and a second lentivirus vector encoding a reverse tetracycline\u2010controlled transactivator (rtTA) that activates transgene expression from the TRE in the presence of DOX.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We evaluated a number of different rtTAs and found rtTA2<jats:sup>S<\/jats:sup>\u2010M2 to induce the highest levels of transgene expression. Regulated transgene expression was stable in human breast carcinoma cells implanted into nude mice for up to 11 weeks. In an attempt to minimize background expression levels, the chicken \u03b2\u2010globin cHS4 insulator element was cloned into the 3\u2032 long terminal repeat (LTR) of the transgene transfer vector. The cHS4 insulator element reduced background expression but expression levels following DOX addition were lower than those observed with vectors lacking an insulator sequence. In a second strategy, vectors bearing second\u2010generation TREs harboring repositioned tetracycline operator elements were used. Such vectors displayed greatly reduced leakiness in the absence of DOX and induced transgene expression levels were up to 522\u2010fold above those seen in the absence of DOX.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Inducible lentivirus vectors bearing insulators or second\u2010generation TREs will likely prove useful for applications demanding the lowest levels of background expression. Copyright \u00a9 2005 John Wiley &amp; Sons, Ltd.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jgm.712","type":"journal-article","created":{"date-parts":[[2005,1,17]],"date-time":"2005-01-17T17:16:24Z","timestamp":1105982184000},"page":"803-817","source":"Crossref","is-referenced-by-count":88,"title":["Tight control of transgene expression by lentivirus vectors containing second\u2010generation tetracycline\u2010responsive promoters"],"prefix":"10.1002","volume":"7","author":[{"given":"Krzysztof","family":"Pluta","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Milson 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