Ubiquitous expression of the rtTA2S-M2 inducible system in transgenic mice driven by the human hnRNPA2B1/CBX3 CpG island.
BACKGROUND
A sensitive, ubiquitously expressed tetracycline inducible system would be a valuable tool in mouse transgenesis. However, this has been difficult to obtain due to position effects observed at different chromosomal sites of transgene integration, which negatively affect expression in many tissues. The aim of this study was to test the utility of a mammalian methylation-free CpG island to drive ubiquitous expression of the sensitive doxycycline (Dox) inducible rtTA2S-M2 Tet-transactivator in transgenic mice.


RESULTS
An 8 kb genomic fragment from the methylation-free CpG island of the human hnRNPA2B1-CBX3 housekeeping gene locus was tested. In a number of transgenic mouse lines obtained, rtTA2S-M2 expression was detected in many tissues examined. Characterisation of the highest expressing rtTA2S-M2 transgenic mouse line demonstrated Dox-inducible GFP transgene expression in many tissues. Using this line we also show highly sensitive quantitative induction with low doses of Dox of an assayable plasma protein transgene under the control of a Tet Responsive Element (TRE). The utility of this rtTA2S-M2 line for inducible expression in mouse embryos was also demonstrated using a GATA-6 Tet-inducible transgene to show specific phenotypes in the embryonic lung, as well as broader effects resulting from the inducible widespread overexpression of the transgene.


CONCLUSIONS
The ubiquitously expressing rtTA2S-M2 transgenic mouse line described here provides a very useful tool for studying the effects of the widespread, inducible overexpression of genes during embryonic development and in adult mice.
Katsantoni
Eleni Z
EZ
Department of Cell Biology, Erasmus University Medical Center, PO Box 2040, 3000 CA Rotterdam, The Netherlands. ekatsantoni@bioacademy.gr
Anghelescu
Nora E
NE
Rottier
Robbert
R
Moerland
Matthijs
M
Antoniou
Michael
M
de Crom
Rini
R
Grosveld
Frank
F
Strouboulis
John
J
D000818:Animals; D015152:Blotting, Northern; D018899:CpG Islands / Q000235:genetics*; D004318:Doxycycline / Q000494:pharmacology; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D038101:Insulator Elements / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D049294:Phospholipid Transfer Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D019076:Transgenes
D049294:Phospholipid Transfer Proteins; D049452:Green Fluorescent Proteins; D004318:Doxycycline