B6;129S-Nkd1^tm1Kwha/J

Stock number: 006958
Research areas: Cell Biology Research, Research Tools

Description

Because the naked cuticle (NKD) genes are considered targets for Wnt/β-catenin signaling, these nkd1^lacZ mutant mice (as well as the similar nkd2^lacZ mutant strain; Stock No. 006960) may be useful as a reporter for Wnt/β-catenin-dependent transcriptional activity during embryonic development or in adult mice.

Detailed description

Mice homozygous for this nkd1^lacZ mutant allele are viable and fertile with slightly reduced mean litter sizes. Northern blot of lung tissue, as well as western blot of embryonic tissue, confirms that the deleted exons (encoding the Dvl-binding domain) are not expressed in homozygous mutants. However, the endogenous promoter directs expression of the internal ribosome entry site-β-galactosidase (IRES-lacZ) fusion gene in a manner that closely mimics the patterns of the endogenous gene mRNAs. Because the naked cuticle (NKD) genes are considered targets for Wnt/β-catenin signaling, these nkd1^lacZ mutant mice (as well as the similar nkd2^lacZ mutant strain; Stock No. 006960) may be useful as a reporter for Wnt/β-catenin-dependent transcriptional activity during embryonic development or in adult mice.

Development

A targeting vector was designed to insert a stop codon into exon 5 at amino acid 91, as well as replace exons 5-10 of the targeted gene with an internal ribosome entry site-β-galactosidase (IRES-lacZ) fusion gene and neomycin cassette. The construct was electroporated into 129SvEv-derived embryonic stem (ES) cells. Correctly targeted ES cells were microinjected into blastocysts, and chimeric offspring were bred to C57BL/6 mice. Heterozygous mutants were bred to generate homozygotes, which were bred together for many generations prior to arrival at The Jackson Laboratory.

Allele

Symbol: Nkd1^tm1Kwha
Name: targeted mutation 1, Keith Wharton
MGI accession ID: MGI:3714460
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Nkd1
Marker name: naked cuticle 1
Chromosome: 8
Strain of origin: 129/SvEv
Expressed genes: lacZ,beta-galactosidase,E. coli
Site of expression: lacZ expression closely mimics the patterns of the endogenous gene.
Molecular note: A targeting vector was designed to insert a stop codon into exon 5 at amino acid 91, as well as replace exons 5-10 of the targeted gene with an internal ribosome entry site-b-galactosidase (IRES-lacZ) fusion gene and neomycin cassette. The construct was electroporated into 129SvEv-derived embryonic stem (ES) cells. Northern blot analysis of lung tissue and Western blot analysis of embronic tissue in homozygotes confirm lack of normal mRNA expression, while X-gal staining closely mimics dynamic expression pattern of normal Nkd1 mRNA.