C57BL/6J-Ank2^tm1.1Csbd/J

Stock number: 038728
Product name: Ank2^fl
Research areas: Neurobiology Research, Research Tools

Description

Ank2^fl mice have exon 4 floxed. This strain may be useful for studying linker function between cytoskeleton and transmembrane protein in neurons associated with cardiac dysfunctions and the onset of Autism spectrum disorders (ASD) and seizures.

Detailed description

Ankyrin 2, brain (Ank2) encodes a neuronal scaffolding protein involved in cell motility, activation, proliferation, contact and the maintenance of specialized membrane domains. In addition, it plays a role in endocytosis and intracellular protein transport, and as a linker between underlying cytoskeleton and several transmembrane proteins in the heart. It has been associated with cardiac dysfunctions and, more recently, has emerged as one of the most frequently mutated Autism spectrum disorders (ASD)-risk genes.

This strain contains loxP sites flanking exon 4 which encodes the N-terminal region and is located between the multiple ankyrin repeats domain and the a highly conserved spectrin binding domain. Deletion of exon 4 affects both short and long variants.

Mice that are homozygous for this allele are viable and fertile and have no overt phenotype. They express normal ANK2 levels. When bred to a strain expressing Cre recombinase, resulting offspring will have exon 4 removed in the cre-expressing tissues of the offspring.

Global Ank2 KO results in homozygous embryonic lethality.

When bred to Emx1-Cre mice (Stock No. 005628), homozygous Ank2 deletion in cortical and hippocampal excitatory neurons leads to ASD-related behavioral deficits and juvenile seizure-related death. Progeny exhibit increased cortical neuronal excitability and firing through decreases in the level and function of Kv7.2/3 potassium channels.

Development

A targeting vector was designed to contain (from 5' to 3') an frt site, an En2 splice acceptor, an internal ribosome entry site (IRES) fused to an EGFP sequence with a polyadenylation signal, and a floxed cassette containing a neo and a second frt site all upstream of exon 4 of the ankyrin 2 (Ank2) gene. A third loxP was placed downstream of exon 4. The construct was electroporated into C57BL/6J-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting Ank2^fl mice were bred with protamine-cre mice on a C57BL/6 genetic background to remove the frt-flanked cassette and one of the three loxP sites. Mice were further bred to C57BL/6J genetic mice (Stock No. 000664) for at least six generations. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Ank2^tm1.1Csbd
Name: targeted mutation 1, Eunjoon Kim
MGI accession ID: MGI:7537121
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Marker symbol: Ank2
Marker name: ankyrin 2, brain
Marker synonyms: ANK-2; Ank3; ankyrin B; Ankyrin-2; Ankyrin-B; brank-2; CFAP87; FAP87; Gm4392; LQT4
Chromosome: 3
Strain of origin: C57BL/6J
Molecular note: The targeting vector is designed to contain (from 5'; to 3') an FRT site, an En2 splice acceptor, an internal ribosome entry site (IRES) fused to an EGFP sequence with a polyadenylation signal, and a floxed cassette containing a neo and a second FRT site all upstream of exon 4. A third loxP was placed downstream of exon 4. The frt-flanked neomycin cassette was removed in the germline via FLP-mediated recombination.