STOCK H2az2^{Tg(Wnt1-cre)11Rth} Tg(Wnt1-GAL4)11Rth/J

Stock number: 003829
Product name: Wnt-1/GAL4/cre-11
Research areas: Developmental Biology Research, Research Tools

Description

These Wnt-1/GAL4/cre-11 transgenic mice have expression of both Cre recombinase and the GAL4 transcriptional activator directed to midbrain and developing neural tube by the wingless-related MMTV integration site 1 (Wnt1) promoter/regulatory sequences. When Wnt-1/GAL4/cre-11 transgenic mice are bred to mice containing loxP site-flanked sequences, cre-mediated recombination results in deletion of the floxed sequences in the midbrain and developing neural tube of the resulting offspring.

Wnt-1/GAL4/cre-11 mice exhibit transgenic overexpression of Wnt1, both within and outside of the expected pattern of expression, which results in increased activation of WNT signaling, along with midbrain enlargement and accompanying loss of midbrain dopaminergic neurons (Lewis et al. 2013 Dev Biol 379:229). Further, it was reported in Goodwin et al. 2019 Genome Res. 29:494, that the Wnt1-cre transgene integrated into chromosome 11 causing a 31,283 bp deletion in the H2afv locus. In Stock No. 009107, founder line 1 had a copy number of greater than 5.

Detailed description

When homozygous for both co-injected transgenes, Wnt-1/GAL4/cre-11 transgenic mice are viable and fertile. Both Cre recombinase and the GAL4 transcriptional activator are expressed under the direction of wingless-related MMTV integration site 1 (Wnt1) promoter/regulatory sequences. Cre recombinase activity is detected in the Wnt1 pattern of expression: in the midbrain by 8.5 dpc and, after neural tube closure, in the dorsal and ventral midlines of the midbrain and caudal diencephalon, midbrain-hindbrain junction and dorsal spinal cord. Double transgenic mice exhibit psychiatric disorder-related behavioral abnormalities. Males, specifically, exhibit increased locomotor activity, increased anxiety, and decreased social interaction, while females exhibit impaired short-term spatial memory and nesting behavior. Both sexes have disordered cholinergic and glutamatergic fiber tracts from the medial habenula neurons in the interpeduncular nucleus.

Wnt-1/GAL4/cre-11 mice exhibit transgenic overexpression of Wnt1, both within and outside of the expected pattern of expression, which results in increased activation of WNT signaling, along with midbrain enlargement and accompanying loss of midbrain dopaminergic neurons (Lewis et al. 2013 Dev Biol 379:229).

When Wnt-1/GAL4/cre-11 transgenic mice are bred to mice containing loxP site-flanked sequences, cre-mediated recombination results in deletion of the floxed sequences in the midbrain and developing neural tube of the resulting offspring.

Development

The transgenic line Wnt-1/GAL4/cre-11 was created by Dr. David H. Rowitch and Andrew P. McMahon (Harvard University). These mice resulted from co-injection of both the pWEXP2-GAL4 transgene (Wnt1-GAL4) and the pWEXP3C-cre transgene (Wnt1-Cre) into pronuclei of B6CBAF1/J (C57BL/6JxCBA/J) zygotes. Double transgenic mice from founder line 11 were obtained and bred with Swiss albino outbred mice. Double transgenic albino mice were sent to The Jackson Laboratory Repository in 2000 (as Stock No. 003829). Upon arrival, mice were bred together to establish the colony. The description of each transgene is below.

The pWEXP2-GAL4 transgene (Wnt1-GAL4) was designed by Dr. David H. Rowitch and Andrew P. McMahon (Harvard University) with the mouse Wnt1 promoter/enhancer elements upstream of a sequence encoding the full-length yeast GAL4 gene. Specifically, the Wnt1-GAL4 transgene was created by inserting a sequence encoding the full-length yeast GAL4 gene into a 10 kbp modified mouse Wnt1 genomic sequence at a polylinker site between the promoter and enhancer (the Wnt1 genomic sequence spans from approximately 1 kbp upstream of the translation initiation codon to approximately 5.5 kbp downstream of the polyA signal, and is modified with a polylinker that disrupts the translational start site and a reverse-oriented neo cassette in 3' UTR of exon 4).

The pWEXP3C-cre transgene (Wnt1-Cre) was designed by Dr. David H. Rowitch and Andrew P. McMahon (Harvard University) with the mouse Wnt1 promoter/enhancer elements upstream of a Cre recombinase cDNA sequence. Specifically, the Wnt1-Cre transgene was created by inserting the Cre recombinase cDNA into a 10 kbp modified mouse Wnt1 genomic sequence at a polylinker site between the promoter and enhancer (the Wnt1 genomic sequence spans from approximately 1 kbp upstream of the translation initiation codon to approximately 5.5 kbp downstream of the polyA signal, and is modified with a polylinker that disrupts the translational start site and a reverse-oriented neo cassette in 3' UTR of exon 4).
In Goodwin et al. 2019 Genome Res. 29:494, it was discovered that the Wnt1-cre transgene integrated into chromosome 11 causing a 31,283 bp deletion in the H2afv locus. In Stock No. 009107, founder line 1 had a copy number of greater than 5.

Allele

Symbol: Tg(Wnt1-GAL4)11Rth
Name: transgene insertion 11, David H Rowitch
MGI accession ID: MGI:3524966
Generation method: Transgenic
Attributes: Inserted expressed sequence
Marker symbol: Tg(Wnt1-GAL4)11Rth
Marker name: transgene insertion 11, David H Rowitch
Chromosome: UN
Strain of origin: (C57BL/6J x CBA/J)F1
Expressed genes: GAL4,transcriptional activator GAL4,yeast
Site of expression: embryonic neural tube, midbrain, dorsal and ventral midlines of the midbrain and caudal diencephalon, the mid-hindbrain junction and dorsal spinal cord
Molecular note: The transgene contains the yeast GAL4 gene directed by Wnt1 promoter/enhancer sequences. Expression of the transgene matches that of the endogenous Wnt1.
General note: Six lines were generated.

Homozygous transgenic mice that are also homozygous for Tg(Wnt1-cre)11Rth are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. Both Cre recombinase and the GAL4 transcriptional activator are expressed under the direction of Wnt1 regulatory sequences. Regulated expression initially occurs in the midbrain. After neural tube closure, expression occurs in the dorsal and ventral midlines of the midbrain and caudal diencephalon, the midbrain-hindbrain junction, and in the dorsal spinal cord.

Allele

Symbol: H2az2^{Tg(Wnt1-cre)11Rth}
Name: transgene insertion 11, David H Rowitch
MGI accession ID: MGI:2386570
Generation method: Transgenic
Attributes: Recombinase-expressing
Marker symbol: H2az2
Marker name: H2A.Z histone variant 2
Chromosome: 11
Strain of origin: (C57BL/6J x CBA/J)F1/J
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: embryonic neural tube, midbrain, dorsal and ventral midlines of the midbrain and caudal diencephalon, the mid-hindbrain junction and dorsal spinal cord
Molecular note: The Wnt1 promoter preceded the cre recombinase coding sequence which was followed downstream by the Wnt1 enhancer. The Wnt1 regulatory sequences initially direct expression in the midbrain. After neural tube closure, expression occurs in the dorsal and ventral midlines of the midbrain and caudal diencephalon, the midbrain-hindbrain junction and in the dorsal spinal cord. Line 11 inserted into the gene at 6425500-6456783 (Build GRCm38/mm10) resulting in a 31,283 bp deletion. Founder line x has a copy number of greater than 5.
General note: Homozygous transgenic mice that are also homozygous for Tg(Wnt1-GAL4)11Rth are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities.