biostudies-literatureUnknown20(1)Taguchi JTemporal control of transgenes has advanced biomedical interventions, including in vivo reprogramming, often utilizing the doxycycline (Dox)-mediated Tet-ON system. Here, we developed the Dox-mediated Tet-ON or complementary Tet-OFF counterpart to thoroughly investigate spatial and temporal transgene regulation in adult tissues, revealing inherent limitations and unexpected capabilities of each system. In stark contrast with the Tet-ON system, which was effective only in particular tissues and cell types, primarily epithelial cells, the Tet-OFF system proved capable of gene induction across diverse cell types. Despite the drawback of the Tet-OFF system in inducibility and tunability identified in our study, we demonstrated that use of tetracycline (Tc) effectively addresses these issues, pStem cell reports102373https://www.ebi.ac.uk/biostudies/studies/S-EPMC11784451biostudies-literatureA versatile in vivo platform for reversible control of transgene expression in adult tissues.PMC11784451Taguchi JOzawa MOhta SYamamoto TYamada YNakasuka FfalseA versatile in vivo platform for reversible control of transgene expression in adult tissues.Temporal control of transgenes has advanced biomedical interventions, including in vivo reprogramming, often utilizing the doxycycline (Dox)-mediated Tet-ON system. Here, we developed the Dox-mediated Tet-ON or complementary Tet-OFF counterpart to thoroughly investigate spatial and temporal transgene regulation in adult tissues, revealing inherent limitations and unexpected capabilities of each system. In stark contrast with the Tet-ON system, which was effective only in particular tissues and cell types, primarily epithelial cells, the Tet-OFF system proved capable of gene induction across diverse cell types. Despite the drawback of the Tet-OFF system in inducibility and tunability identified in our study, we demonstrated that use of tetracycline (Tc) effectively addresses these issues, p2025-01-01T00:00:00Z2025 Jan2026-06-01T12:56:05.471Z2025-04-04T01:13:19.778ZS-EPMC117844513964287410.1016/j.stemcr.2024.11.003