B6;129-Gt(ROSA)26Sor^{tm3(rtTA,tetO-cre/ERT)Nat}/J

Stock number: 010557
Research areas: Research Tools

Description

These R26rtTACreER mice have a CreER^T, reverse tetracycline trans-activator (rtTA), and Tet-responsive element (TRE) targeted to the mouse Gt(ROSA)26Sor gene to create a gene expression tool with two independent layers of pharmacologic control and a widespread pattern of activity. In this construct, expression of tamoxifen-sensitive CreER^T is under the control of rtTA. Upon administration of doxycycline, rtTA activates transcription of the CreER coding region. This enhances the dynamic range of CreER and enables sparse labeling in situations where reduced recombination efficiency facilitates behavioral, morphological or functional studies of modified cells among populations of unmodified neighbors.

Detailed description

These R26rtTACreER mice have a CreER^T, reverse tetracycline trans-activator (rtTA), and Tet-responsive element (TRE) targeted to the mouse Gt(ROSA)26Sor gene to create a gene expression tool with two independent layers of pharmacologic control and a widespread pattern of activity. In this construct, expression of tamoxifen-sensitive CreER^T is under the control of rtTA, a sequence-specific DNA-binding protein that activates transcription upon binding tetracycline derivatives such as doxycycline. Upon administration of doxycycline, rtTA activates transcription of the CreER coding region by binding to the Tet operator located immediately 5' of a minimal promoter. This enhances the dynamic range of CreER and enables sparse labeling in situations where reduced recombination efficiency facilitates behavioral, morphological or functional studies of modified cells among populations of unmodified neighbors.

Development

A splice acceptor, reverse tetracycline-responsive transcriptional activator (rtTA), six repeats of an SV40 transcriptional terminator, tetracycline response element (TRE) enhancer-promoter, CreER fusion protein open reading frame, bovine growth hormone 3' UTR and transcriptional terminator region were introuduced to exon 1 of the targeted gene. The vector was introduced to 129-derived embryonic stem cells. This line was backcrossed 3 times to C57BL/6 by the donating laboratory.

Allele

Symbol: Gt(ROSA)26Sor^{tm3(rtTA,tetO-cre/ERT)Nat}
Name: targeted mutation 3, Jeremy Nathans
MGI accession ID: MGI:3849063
Generation method: Targeted
Attributes: Recombinase-expressing; Inducible; Transactivator
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: 129
Expressed genes: cre/ER,cre inducible estrogen receptor,bacteriophage P1/Human; rtTA,reverse tetracycline-controlled transactivator,E. coli
Site of expression: widespread, doxycycline inducible expression of Cre recombinase
Molecular note: The construct consists of the rtTA coding region preceded by a splice acceptor so that it can be spliced onto the 5' exon (exon1) of the endogenous ROSA26 transcript, and is followed by six repeats of an SV40 transcriptional terminator signal ("stop"). In the presence of doxycycline, the rtTA protein activates transcription from the seven head-to-tail Tet response elements immediately 5' of a minimal cytomegalovirus immediate early promoter (TRE) of the CreER fusion protein open reading frame (followed by the bovine growth hormone 3' UTR and transcriptional terminator region), leading to expression of CreER.