C57BL/6NJ-Kcnt1^em8Frk/J

Stock number: 028830
Product name: Kcnt1-Y777H
Strain synonyms: Kcnt1 Y777H KI
Research areas: Cell Biology Research, Mouse/Human Gene Homologs, Neurobiology Research

Description

This CRISPR/Cas9 generated mutant of the Kcnt1, potassium channel, subfamily T, member 1, gene carries a Y777H point mutation.

Detailed description

CRISPR/Cas9 endonuclease mediated genome editing of the Kcnt1, potassium channel, subfamily T, member 1, gene was used to introduce a Y777H point mutation. The targeted Kcnt1 gene (also known as KCa4.1 and Slo2.2) encodes a subunit of a sodium activated potassium channel. Mutations in this gene have been associated with Early Infantile Epileptic Encephalopathy and Fragile X syndrome. Homozygous Kcnt1-Y777H mice have frequent, sleep-associated, spontaneous seizures (Shore et al. 2020 Cell Rep. 33:108303 [PMID:33113364]).

Development

Plasmids encoding a signal guide RNA designed to introduce a TAT->CAT point mutation at amino acid 777 (Y777H) in the Kcnt1 gene and the cas9 nuclease were introduced into the cytoplasm C57BL/6NJ-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further bred to C57BL/6NJ (Stock No. 005304) to develop the colony.
Please note, the citation for this Kcnt1^em8Frk allele is Shore et al. 2020 Cell Rep. 33:108303 [PMID:33113364], which refers to it as Kcnt1^{em1(Y777H)Frk}.

Allele

Symbol: Kcnt1^em8Frk
Name: endonuclease-mediated mutation 8, Wayne N Frankel
MGI accession ID: MGI:6509315
Generation method: Endonuclease-mediated
Attributes: Not Specified
Synonyms: Kcnt1^{em1(Y777H)Frk}; Kcnt1^em1Frk; Kcnt1^m; Kcnt1^Y777H
Marker symbol: Kcnt1
Marker name: potassium channel, subfamily T, member 1
Marker synonyms: bA100C15.2; C030030G16Rik; DEE14; EIEE14; ENFL5; KCa4.1; rSlo2; Slack; slo2; Slo2.2
Chromosome: 2
Strain of origin: C57BL/6NJ
Molecular note: Using sgRNAs and an ssODN template with CRISPR/cas9 technology, a T-to-C point mutation (c.2329T>C) was created to change tyrosine codon 777 (TAT) to a histidine codon (CAT) (p.Y777H). This mutation (formerly designated em1) mimics the p.Y796H gain-of-function mutation found in human autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) patients.