B6.129P2-Pdha1^tm1Ptl/J

Stock number: 017443
Research areas: Diabetes and Obesity Research, Internal/Organ Research, Metabolism Research, Research Tools

Description

These Pdha1^flox8 mutant mice possess loxP sites flanking exon 8 of the pyruvate dehydrogenase E1 alpha 1 (Pdha1) gene. This strain may be useful for studying lipid metabolism, glucose metabolism, and insulin sensitivity.

Detailed description

These Pdha1^flox8 mutant mice possess loxP sites flanking exon 8 of the pyruvate dehydrogenase E1 alpha 1 (Pdha1) gene. Mice that are homozygous for this allele are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. PDHA1 is catalytic component of the pyruvate dehydrogenase complex (PDC) which is a mitochondrial multienzyme complex involved in lipid synthesis, glucose homeostasis, and metabolism of carbohydrates. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 8 deleted in cre-expressing tissues. For example, when crossed to a strain expressing Cre recombinase in the liver, this mutant mouse strain may be useful for studying lipid metabolism, glucose metabolism, and insulin sensitivity in the liver.

When bred to a strain expressing Cre recombinase in the central and periphal nervous system (see Stock No. 003771 for example), this mutant mouse strain may be useful in studies of neuronal migration, axonal growth and cell-cell interactions.

When bred to a strain expressing Cre recombinase in the oocyte (see Stock No. 003651 for example), this mutant mouse strain may be useful in studies of oogenesis.

When bred to a strain expressing Cre recombinase in skeletal and cardiac muscle (see Stock No. 006475 for example), this mutant mouse strain may be useful in studies of cardiac glucose metabolism.

Development

A targeting vector was designed to insert a single loxP site upstream of exon 8, and a loxP-flanked neomycin resistance (neo) cassette downstream of exon 8 of the pyruvate dehydrogenase E1 alpha 1 (Pdha1) gene. The construct was electroporated into 129P2/OlaHsd-derived E14.1 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a pMC-Cre expression plasmid to delete the neo cassette. Resulting ES cells contained multiple gene rearrangments; intact floxed-exon 8, intact floxed-neo cassette, or excision of both exon 8 and the neo cassette. Correctly targeted ES cells, containing only the floxed-exon 8, were injected into C57BL/6 blastocysts and resulting chimeric males were bred with 129 females. These mice were backcrossed to C57BL/6 mice for at least 10 generations to establish a colony of Pdha1^flox8 mice. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation.

Allele

Symbol: Pdha1^tm1Ptl
Name: targeted mutation 1, Mulchand S Patel
MGI accession ID: MGI:2386703
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Pdha1
Marker name: pyruvate dehydrogenase E1 alpha 1
Chromosome: X
Strain of origin: 129P2/OlaHsd
Molecular note: Exon 8 was left flanked by loxP sites after a downstream floxed tk-neo cassette was excised by the in vitro expression of cre recombinase. Western blot analysis of homozygous mutant ES cells showed normal levels of both isoforms produced by the endogenous allele. Mice carrying this allele were bred to transgenic mice expressing cre in the germline to generate Pdha1^tm1.1Ptl.