A potentially immunologically inert derivative of the reverse tetracycline-controlled transactivator.
The archetypical system for regulating heterologous gene expression in mammalian cells involves tetracycline-activated transactivators (rtTA). Binding of such transactivators to tet-operator-controlled promoters induces transcription. Immune responses directed against the transactivator proteins may limit the applicability of this system in immune-competent hosts. To circumvent such immune responses the immune evasion mechanism of the Epstein-Barr virus Nuclear-Antigen 1 was exploited. Our data show that fusion of the rtTA with the EBNA-1 derived Gly-Ala repeat yielded an efficient transactivator with no detectable activity in absence of inducer. Antigenic peptides of the fusion protein were not presented in MHC class I.
Zaldumbide
Arnaud
A
Department of Molecular Cell Biology, Leiden University Medical Center, Postal Zone S1-P, P.O. Box 9600, 2300 RC, Leiden, The Netherlands. a.zaldumbide@lumc.nl
Weening
Susan
S
Cramer
Steve J
SJ
Rabelink
Martijn J W E
MJ
Verhaagen
Joost
J
Hoeben
Rob C
RC
D017951:Antigen Presentation; D002460:Cell Line; D019309:Epstein-Barr Virus Nuclear Antigens / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression; D004854:Herpesvirus 4, Human / Q000235:genetics; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D006801:Humans; D057131:Immune Evasion; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013752:Tetracycline / Q000378:metabolism*; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology
D019309:Epstein-Barr Virus Nuclear Antigens; D015395:Histocompatibility Antigens Class I; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D013752:Tetracycline; C417029:EBV-encoded nuclear antigen 1