This CRISPR/Cas9 generated mutant of the Kcnt1, potassium channel, subfamily T, member 1, gene carries a Y777H point mutation.
Wayne Frankel, Columbia University
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]).
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 Kcnt1em8Frk allele is Shore et al. 2020 Cell Rep. 33:108303 [PMID:33113364], which refers to it as Kcnt1em1(Y777H)Frk.
|Allele Name||endonuclease-mediated mutation 8, Wayne N Frankel|
|Allele Type||Endonuclease-mediated (Not Specified)|
|Allele Synonym(s)||Kcnt1em1(Y777H)Frk; Kcnt1em1Frk; Kcnt1m; Kcnt1Y777H|
|Gene Symbol and Name||Kcnt1, potassium channel, subfamily T, member 1|
|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.|
Heterozygotes may be bred to C57BL/6NJ (Stock No. 005304) or wildtype littermates. Homozygous Kcnt1-Y777H mice have frequent, sleep-associated, spontaneous seizures.
When using the Kcnt1-Y777H mouse strain in a publication, please cite the originating article(s) and include JAX stock #028830 in your Materials and Methods section.