title: Silencing and un-silencing of tetracycline-controlled genes in neurons.
abstract: To identify the underlying reason for the controversial performance of tetracycline (Tet)-controlled regulated gene expression in mammalian neurons, we investigated each of the three components that comprise the Tet inducible systems, namely tetracyclines as inducers, tetracycline-transactivator (tTA) and reverse tTA (rtTA), and tTA-responsive promoters (P(tets)). We have discovered that stably integrated P(tet) becomes functionally silenced in the majority of neurons when it is inactive during development. P(tet) silencing can be avoided when it is either not integrated in the genome or stably-integrated with basal activity. Moreover, long-term, high transactivator levels in neurons can often overcome integration-induced P(tet) gene silencing, possibly by inducing promoter accessibility.
authors.affiliation: Max Planck Institute for Medical Research, Heidelberg, Germany.
authors.forename: Peixin
authors.initials: P
authors.lastname: Zhu
authors.forename: M Isabel
authors.initials: MI
authors.lastname: Aller
authors.forename: Udo
authors.initials: U
authors.lastname: Baron
authors.forename: Sidney
authors.initials: S
authors.lastname: Cambridge
authors.forename: Melanie
authors.initials: M
authors.lastname: Bausen
authors.forename: Jan
authors.initials: J
authors.lastname: Herb
authors.forename: Jürgen
authors.initials: J
authors.lastname: Sawinski
authors.forename: Ali
authors.initials: A
authors.lastname: Cetin
authors.forename: Pavel
authors.initials: P
authors.lastname: Osten
authors.forename: Mark L
authors.initials: ML
authors.lastname: Nelson
authors.forename: Sebastian
authors.initials: S
authors.lastname: Kügler
authors.forename: Peter H
authors.initials: PH
authors.lastname: Seeburg
authors.forename: Rolf
authors.initials: R
authors.lastname: Sprengel
authors.forename: Mazahir T
authors.initials: MT
authors.lastname: Hasan
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics*; D005786:Gene Expression Regulation; D020868:Gene Silencing / Q000187:drug effects*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D017403:In Situ Hybridization; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*; D015534:Trans-Activators / Q000187:drug effects / Q000235:genetics*; D015533:Transcriptional Activation / Q000187:drug effects
chemical_list: D015534:Trans-Activators; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008156:Luciferases; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D013752:Tetracycline