B6;129S-Pkm^tm1.1Mgvh/J

Stock number: 024048
Product name: Pkm2^fl
Research areas: Cancer Research, Research Tools

Description

These Pkm2^fl/fl floxed mutant mice possess loxP sites flanking exon 10 of the Pkm gene. This strain may be useful for generating conditional mutations in applications related to the role of pyruvate kinase M2 isoform in tumor formation.

Detailed description

The pyruvate kinase muscle (Pkm) gene encodes different splicing variant isoenzymes which are involved in glycolysis. This gene produces an RNA product that is alternatively spliced to generate mRNAs encoding two different isoforms of the enzyme: PKM1 and PKM2. The mRNA encoding PKM1 and PKM2 differ only in inclusion of either exon 9 for the PKM1 message or exon 10 for the PKM2 message. The pyruvate kinase M2 isoform is found in tumor cells and some normal tissues. These Pkm2^fl/fl mice possess loxP sites on either side of exon 10 of the targeted gene. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 10 deleted in the cre-expressing tissues: pyruvate kinase M2 isoform expression is eliminated, while pyruvate kinase M1 isoform expression is unchanged.

When bred to a cancer model, this mutant mouse strain may be useful in studies of acceleration of tumorigenesis and metastasis. It could also be used to test the role of PKM2 in normal tissue physiology.

Development

A targeting vector containing a FRT site flanked NEO cassette upstream of exon 9 of the targeted gene, and loxP sites flanking exon 10 was electroporated into 129S6/SvEvTac derived TC1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were crossed to mice (on an unconfirmed B6;129 genetic background) expressing FLP1 (FlpO) recombinase (ROSA26Flpo) to excise the NEO cassette. Pkm2^fl/fl mice that retained the loxP site flanked exon 10 were then bred to remove the FLP1 (FlpO) recombinase allele. Heterozygotes were crossed to generate homozygotes. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Pkm^tm1.1Mgvh
Name: targeted mutation 1.1, Matthew G Vander Heiden
MGI accession ID: MGI:5547750
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Modified isoform(s); No functional change
Synonyms: Pkm2^fl
Marker symbol: Pkm
Marker name: pyruvate kinase, muscle
Marker synonyms: CTHBP; HEL-S-30; OIP3; p58; PK; Pk-2; PK3; Pk-3; PKM12; Pkm2; TCB; THBP1
Chromosome: 9
Strain of origin: 129S6/SvEvTac
Molecular note: A targeting vector was designed to contain an FRT-flanked neomycin cassette upstream of exon 9, and loxP sites that flank exon 10 which is alternatively spliced only in the Pkm2 isoform. Mutant mice were bred with mice expressing FlpO recombinase (Gt(ROSA)26Sor) to remove the neomycin selection cassette. When these mutant mice are bred to mice that express Cre recombinase, the resulting offspring will have exon 10 deleted in the cre-expressing tissues: pyruvate kinase M2 isoform expression is eliminated, while pyruvate kinase M1 isoform expression is unchanged.