B6;CBA-Axin2^em1Fstl/J

Stock number: 031387
Product name: Axin2-TQ
Research areas: Cell Biology Research, Neurobiology Research, Research Tools

Description

Axin2-TQ is a knock-in/knock-out allele expressing the mTurquoise2 fluorescent reporter from the Axin2 locus. These mice are suitable for use in applications related to the detection of canonical Wnt signaling in living cells.

Detailed description

CRISPR/cas9 endonuclease mediated genome editing of the axin 2 (Axin2) gene was used to introduce an mTurquoise2 reporter knock-in/knock-out mutation. AXIN2 plays an integral role in the Wnt/β-catenin signaling pathway that is involved in stem cell maintenance. As such, it is a frequently-used marker of functional stem cells and a target for analyzing the pathway in a variety of tissues. The Axin2-TQ knock-in/knock-out allele has two copies of the mTurquoise2 reporter replacing the start codon in exon 2 of the Axin2 gene. Homozygous mice are viable and fertile. Wnt signals are detected in the intestinal crypts by direct fluorescent microscopy and in the thymus by flow cytometry. The reporter expression pattern is similar to the endogenous Axin2 gene expression pattern. Homozygous mice lack AXIN2 expression and exhibit curly tails and growth retardation. These mice are suitable for use in applications related to the detection of canonical Wnt signaling in living cells.

Of note, Axin2-mGFP mice (Stock No. 037313) are a knock-in/knock-out mouse line expressing a membrane targeted GFP fluorescent reporter from the Axin2 locus.

Development

Plasmids encoding a signal guide RNA, designed to replace most of exon 2 of the axin 2 (Axin2) gene with an mTurquoise2 (green fluorescent protein in far blue) coding sequence and a frt-flanked neomycin resistance (neo) cassette, and the cas9 nuclease were microinjected into the pronucleus of fertilized oocytes obtained from superovulated B6CBAF1/J donors mated with C57BL6/Jico studs. Exon 2 contains the start codon of the Axin2 gene. The resulting chimeric animals, containing two copies of the reporter cassette, were crossed to C57BL/6 mice. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Axin2^em1Fstl
Name: endonuclease-mediated mutation 1, Frank Staal
MGI accession ID: MGI:6111808
Generation method: Endonuclease-mediated
Attributes: Reporter; Null/Knockout
Synonyms: Axin2-TQ
Marker symbol: Axin2
Marker name: axin 2
Marker synonyms: Axil; axin-2; Conductin; ODCRCS
Chromosome: 11
Strain of origin: (C57BL/6J x CBA/J)F1 x C57BL/6JIcoCrlLumc
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: mTurquoise2 fluorescent protein expression pattern is similar to the endogenous Axin2 gene expression pattern with Wnt signals detected in the intestinal crypts by direct fluorescent microscopy and in the thymus by flow cytometry...
Molecular note: CRISPR/Cas9 mediated mutatgenesis was used to replace most of exon 2 (including the start codon) with an mTurquoise2 (green fluorescent protein in far blue) coding sequence and a loxP-flanked neomycin resistance (neo) cassette. Reporter expression is under the control of endogenous gene promoter.