C57BL/6J-Ppara^em1Ngwu/J

Stock number: 037968
Product name: PPARα HA knock-in
Research areas: Cell Biology Research, Metabolism Research, Research Tools

Description

PPARα HA knock-in mice express an N-terminal HA-tag under direction of the endogenous Ppara promoter. This mouse strain could be useful for studying the regulatory mechanisms required to drive the energy-burning characteristics of brown adipocytes to counter obesity.

Detailed description

PPARα HA knock-in mice express an N-terminal HA-tag under direction of the endogenous peroxisome proliferator activated receptor alpha (Ppara) promoter. Inserted just downstream of the Ppara start codon, this insertion did not disrupt its endogenous transcriptional regulations. Ppara encodes a nuclear transcription factor that acts as a regulator of lipid metabolism. These mice are useful for immunoprecipitation experiments followed by western blot when studying the energy-burning characteristics of brown adipose tissue (BAT), a tissue specialized for thermogenesis. Upon cold stimulation, brown adipocytes catabolize different sources of carbon-carbon bonds, including self-stored fat, to increase heat generation through the mitochondrial proton gradient. Mice may be helpful for dissecting regulatory mechanisms needed to drive the activation of brown adipocytes to counter obesity. Homozygous mice are viable and fertile.

Development

crRNA/tracrRNA duplex gRNA was designed to insert an N-terminal HA tag immediately downstream of the start codon of the peroxisome proliferator activated receptor alpha (Ppara) gene. The crRNA/tracrRNA, targeting vector, and the Cas9 nuclease protein were introduced into the pronuclei of fertilized eggs from C57BL/6J mice. Targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by PCR and sequencing which also confirmed the absence of random insertions around the insertion site. Resulting PPARα HA knock-in mice were further backcrossed to C57BL/6J mice (Stock No. 000664) for five generations by the donating laboratory. Cryopreserved sperm was sent to The Jackson Laboratory. Upon arrival, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes to establish our live colony.

Allele

Symbol: Ppara^em1Ngwu
Name: endonuclease-mediated mutation 1, Ning Wu
MGI accession ID: MGI:7424957
Generation method: Endonuclease-mediated
Attributes: Epitope tag
Synonyms: PPARalpha HA KI
Marker symbol: Ppara
Marker name: peroxisome proliferator activated receptor alpha
Marker synonyms: 4933429D07Rik; hPPAR; Nr1c1; Ppar; PPARalpha; PPAR-alpha
Chromosome: 15
Strain of origin: C57BL/6J
Molecular note: : crRNA/tracrRNA duplex gRNA is designed to insert an N-terminal HA tag immediately downstream of the start codon.