C57BL/6N-Krt20^{em1(cre/ERT2)Amc}/J

Stock number: 030600
Product name: Krt20-T2A-CRE-ERT2
Research areas: Research Tools

Description

This CRISPR/Cas9 generated mutant of the Krt20 gene carries a cre/ERT2 knock-in mutation immediately before the stop codon.

Detailed description

CRISPR/cas9 endonuclease mediated genome editing of the Krt20, transient receptor potential cation channel, subfamily V, member 4, gene was used to introduce a cre/ERT2 knock-in mutation immediately before the stop codon. The targeted Krt20 gene encodes a type I, acidic, intermediate filament in the gastric and intestinal mucosa, and that is often upregulated in cancer cells. Heterozygotes are viable and fertile. Homozygous viability/fertility has not been tested (May 2017). As the mice are characterized, we will modify the strain description and add phenotype data. When bred to a tdTomato reporter line, the resulting mice, at 3 weeks of age, exhibit tamoxifen inducible tdTomato positive cells in the bladder and gut.

This strain was generated in collaboration with the Genitourinary Development Molecular Anatomy Project (GUDMAP) consortium.

Development

Plasmids encoding a signal guide RNA designed to introduce a cre/ERT2 knock-in mutation (T2A-CRE-ERT2-bGHpA) immediately before the stop codon of the Krt20 gene and the cas9 nuclease were introduced into the cytoplasm of C57BL/6N-derived JM8.N4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into B6(Cg)-Tyr^c-2J/J blastocysts. Correctly targeted pups were identified by sequencing and PCR. The mice were bred to C57BL/6NJ (Stock No. 005304) for 2 generations.

Allele

Symbol: Krt20^{em1(cre/ERT2)Amc}
Name: endonuclease-mediated mutation 1, Andrew P McMahon
MGI accession ID: MGI:5903002
Generation method: Endonuclease-mediated
Attributes: Recombinase-expressing; Inducible
Synonyms: Krt20-T2A-CRE-ERT2
Marker symbol: Krt20
Marker name: keratin 20
Marker synonyms: 9030623C06Rik; CD20; CK20; CK-20; cytokeratin 20; K20; Krt21
Chromosome: 11
Strain of origin: C57BL/6N
Expressed genes: cre/ERT2,Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of expression: When these mice are bred to a reporter line, the resulting mice exhibit tamoxifen inducible fluorescent cells in the bladder and gut.
Molecular note: Plasmids encoding a signal guide RNA designed to introduce a cre/ERT2 knock-in mutation into immediately before the stop codon of the Krt20 gene and the cas9 nuclease were introduced into the cytoplasm of C57BL/6N-derived JM8.N4 ES cells.