129S-Kcnab2^tm2Mes/NystJ

Stock number: 037823
Product name: Kvβ2-Y90F
Research areas: Cell Biology Research, Neurobiology Research

Description

Kvβ2-Y90F knock-in mice have a tyrosine to phenylalanine substitution at position 90 (Y90F). of the Kcnab2 (potassium voltage-gated channel, shaker-related subfamily, beta member 2) gene. This strain may be useful for studies involving potassium channel regulation.

Detailed description

Kcnab2 (potassium voltage-gated channel, shaker-related subfamily, beta member 2) encodes a member of aldo-ketoreductase (AKR) superfamily and is involved in the regulation of voltage-gated potassium currents. Kvβ proteins are expressed in multiple tissues, including the coronary vasculature, in contrast to Kvβ1.1, Kvβ2 functions to enhance Kv-mediated oxygen-dependent coronary vasodilation. Kvβ2-Y90F knockin mice have a tyrosine to phenylalanine substitution at position 90 (Y90F). The mutation results in the abolishment of AKR activity. Mice homozygous for the mutation are viable and fertile with no overt phenotype.

Of note, the knockout version of this allele is distributed as 129S-Kcnab2^tm1Kmc/NystJ, Stock No. 037822.

Development

The Kvβ2-knockin allele was created by Dr. Albert Messing and Dr. Ken McCormack (University of Wisconsin).  The targeting vector was designed to insert a tyrosine to phenylalanine at position 90 (p.Y90F), deletion of the AccI site, and insertion of a SacII site mutation (GTC TTC GCC GCG GGA Aagtacgta) in exon 5 followed by a loxP-flanked neo cassette in intron 5 of the Kcnab2 (potassium voltage-gated channel, shaker-related subfamily, beta member 2) gene on chromosome 4. The construct was electroporated into 129S7/SvEvBrd -derived AB1 embryonic stem (ES) cells. Transient transfection of a CMV-Cre recombinase excised the neo cassette. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred with C57BL/6 females and maintained on a mixed background. Dr. Matthew Nystoriak transferred mice to the University of Louisville. Mice were rederived using 129S/SvEv and backcrossed for 3 generations. To establish our live colony, an aliquot of frozen sperm was used to fertilize 129S1/SvImJ (Stock No. 002488) oocytes.

Allele

Symbol: Kcnab2^tm2Mes
Name: targeted mutation 2, Albee Messing
MGI accession ID: MGI:2179739
Generation method: Targeted
Attributes: Not Specified
Synonyms: Kvbeta2-Y90F
Marker symbol: Kcnab2
Marker name: potassium voltage-gated channel, shaker-related subfamily, beta member 2
Marker synonyms: AKR6A5; F5; HKvbeta2; HKvbeta2.1; HKvbeta2.2; I2rf5; KCNA2B; Kcnb3; KV-BETA-2
Chromosome: 4
Strain of origin: 129S7/SvEvBrd-Hprt1⁺
Molecular note: A tyrosine to phenylalanine substitution at amino acid 90 (p.Y90F) was introduced via homologous recombination. The resulting protein product is enzymatically inactive yet retains structural integrity and DNA binding ability. The Y90F point mutation was verified via RT-PCR and restriction digest analysis of RNA transcripts from mutant animals.