These Wnt9bc mice harbor loxP sites flanking exon 2 of the Wnt9b (wingless-type MMTV integration site 9B) locus and may be useful in studying the role of Wnt9b (and other Wnt family members) in development and canonical Wnt signaling cascades, including metanephric kidney and urogenital system development.
Andrew P McMahon, University of Southern California
Mice homozygous for the Wnt9bc allele are viable and fertile, with loxP sites flanking exon 2 of the targeted gene. When bred to mice that express Cre recombinase, the resulting offspring will have this region deleted in the cre-expressing tissue(s). Such deletion is predicted to result in out-of-frame splicing of exon 1 to exon 3, and consequently a mutant transcript that would encode a nonfunctional peptide comprising the first 27 amino acids of Wnt9b that includes the signal peptide. These Wnt9bc mice may be useful in generating conditional mutations for studying the role of Wnt9b (and other Wnt family members) in development and canonical Wnt signaling cascades, including metanephric kidney and urogenital system development.
A targeting vector was designed to insert a loxP site just upstream of exon 2, and a frt-flanked PGK-Neo cassette followed by a second loxP site just downstream of exon 2 of the targeted gene. This construct was microinjected into 129X1/SvJ-derived AV3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric mice were bred to C57BL/6 inbred mice to generate Wnt9bcneo mice. To remove the frt-flanked selection cassette, Wnt9bcneo mice were then bred to an FLPe-expressing strain (on the Swiss-Webster genetic background). The resulting Wnt9bc offspring (with a loxP site just upstream of exon 2, and a single remaining frt-site followed by a second loxP site just downstream of exon 2) were bred with C57BL/6 mice for approximately 6 generations prior to arrival at The Jackson Laboratory. Upon arrival, these mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony. A SNP (single nucleotide polymorphism) analysis performed by The Jackson Laboratory revealed 17 of 27 markers that were not C57BL/6 allele-type, suggesting this strain is still segregating for Swiss Webster markers.
|Allele Name||targeted mutation 1.2, Andrew P McMahon|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||Wnt9bc; Wnt9bflox|
|Gene Symbol and Name||Wnt9b, wingless-type MMTV integration site family, member 9B|
|Gene Synonym(s)||clf1; Wnt15; cleft lip; cleft lip 1; WNT14B; WNT15; clf; clf1; wingless-type MMTV integration site 15; clf; Wnt15; Wnt14b|
|Strain of Origin||129X1/SvJ|
|Molecular Note||A targeting vector was designed to insert a loxP site just upstream of exon 2, and a frt-flanked PGK-Neo cassette followed by a second loxP site just downstream of exon 2 of the targeted gene to generate Wnt9bMtm1Amc mice (Wnt9bcneo. To remove the frt-flanked selection cassette, these mice were then bred to an FLPe-expressing strain (on the Swiss-Webster genetic background). The resulting Wnt9bc offspring carry a loxP site just upstream of exon 2, and a single remaining frt-site followed by a second loxP site just downstream of exon 2.|
|Mutations Made By|| |
Andrew McMahon, University of Southern California
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