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.
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
Arnaud
A
Zaldumbide
Susan
S
Weening
Steve J
SJ
Cramer
Martijn J W E
MJ
Rabelink
Joost
J
Verhaagen
Rob C
RC
Hoeben
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