This CRISPR-generated knock-out mutant of the thiamine pyrophosphokinase (Tpk1) gene has been generated by the Knockout Mouse Phenotyping Program (KOMP2) at The Jackson Laboratory. Tpk1 encodes an enzyme that catalyzes the phosphorylation of thiamine to thiamine pyrophosphate.Read More +
This strain was generated by the Knockout Mouse Phenotyping Program (KOMP2) at The Jackson Laboratory using CRISPR technology. The targeted gene, thiamine pyrophosphokinase (Tpk1), encodes an enzyme that catalyzes the phosphorylation of thiamine to thiamine pyrophosphate. The alteration resulted in the deletion of 415 bp, which should result in the deletion of exon 4, amino acid change after residue 39, and early termination 49 amino acids later. As mice are characterized, phenotype data will be accessible on the International Mouse Phenotyping Consortium website.
Guide RNAs (CCAGCGAGTCAGGGTCTACA, TAGGAAACCACCTCTGTGAG, TCTCCAGTCCATCTGAACAG and CCAGCTTCGGTAATAAAAGA), designed to delete 415 bp in exon 4 of the thiamine pyrophosphokinase (Tpk1) gene, and Cas9 nuclease were introduced into C57BL/6NJ-derived fertilized eggs with well recognized pronuclei. Embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by PCR and further bred to C57BL/6NJ (Stock No. 005304) to develop the colony.
|Allele Name||endonuclease-mediated mutation 1, Jackson|
|Allele Type||Endonuclease-mediated (Null/Knockout)|
|Gene Symbol and Name||Tpk1, thiamine pyrophosphokinase|
|Strain of Origin||C57BL/6NJ|
|Molecular Note||This allele from project Tpk1-8645J-9675M was generated at The Jackson Laboratory by injecting Cas9 RNA and 4 guide sequences CCAGCGAGTCAGGGTCTACA, TAGGAAACCACCTCTGTGAG, TCTCCAGTCCATCTGAACAG and CCAGCTTCGGTAATAAAAGA, which resulted in a 415 bp deletion beginning at Chromosome 6 negative strand position 43,560,232 bp, CTGTTCAGATGGACTGGAGA, and ending after ACCACCTCTGTGAGAGGGCT at 43,559,818 bp (GRCm38/mm10). This mutation deletes exon 4 and 345 bp of flanking intronic sequence including the splice acceptor and donor. In addition there is a 4 bp deletion (AAAG) 49 bp before the 415 bp deletion that will not alter the results of the exon deletion. This mutation is predicted to cause a change of amino acid sequence after residue 39 and early truncation 49 amino acids later.|
Heterozygotes may be bred to C57BL/6NJ (Stock No. 005304) or wildtype littermates. As mice are characterized, data related to viability and fertility will be provided on the International Mouse Phenotyping Consortium website.
When using the C57BL/6NJ-Tpk1em1(IMPC)J/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #42313 in your Materials and Methods section.