Silencing and un-silencing of tetracycline-controlled genes in neurons.
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.
Zhu
Peixin
P
Max Planck Institute for Medical Research, Heidelberg, Germany.
Aller
M Isabel
MI
Baron
Udo
U
Cambridge
Sidney
S
Bausen
Melanie
M
Herb
Jan
J
Sawinski
Jürgen
J
Cetin
Ali
A
Osten
Pavel
P
Nelson
Mark L
ML
Kügler
Sebastian
S
Seeburg
Peter H
PH
Sprengel
Rolf
R
Hasan
Mazahir T
MT
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
D015534:Trans-Activators; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D008156:Luciferases; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D013752:Tetracycline