Location
Philadelphia, PA
Start Date
8-5-2019 1:00 PM
End Date
8-5-2019 4:00 PM
Description
The global epidemic of obesity and type II diabetes has led to a growing interest in the underlying mechanisms of metabolic diseases. The peroxisome proliferator-activated receptor gamma (PPARγ) is a member of the nuclear receptor superfamily, and is vital for the transcriptional regulation of adipogenesis, insulin sensitivity and lipid metabolism. In the mouse model, it has been demonstrated that global knockout of PPARγ leads to severe metabolic disturbance, resulting in embryonic lethality. However, the specific regulatory roles of its two protein isoforms, PPARγ1 and PPARγ2, remain uncertain, due to limitations of reagents and appropriate mouse models. To investigate the hypothesis that PPARγ1 and PPARγ2 are functionally distinct, we generated PPARγ1 and PPARγ2 tagged mice using CRISPR-Cas9 technology. PPARγ1 and PPARγ2 specific knockout mice were also generated incidentally during this process, via aberrant recombination. By reverse-transcription quantitative PCR (RT-qPCR), and western blot, we confirmed the presence of the appropriate tags in our PPARγ1 and PPARγ2 tagged mice, with no significant disruption to mRNA or protein expression. Furthermore, we found that PPARγ1 mRNA and protein expression levels were reduced in our PPARγ1 knockout model, compared to the wild type. Interestingly, we found that there was a complete loss of PPARγ2 protein expression, despite an increase in PPARγ2 mRNA expression in our PPARγ2 knockout model. These data suggest that we have successfully generated PPARγ1 and PPARγ2 knockin and knockout mice. Our mouse models provide a valuable tool to study the individual roles of PPARγ1 and PPARγ2 in adipogenesis, insulin sensitivity and metabolic disease.
Embargo Period
5-30-2019
Characterization of PPAR-gamma 1 and PPAR-gamma 2 in Knockin and Knockout Mouse Models
Philadelphia, PA
The global epidemic of obesity and type II diabetes has led to a growing interest in the underlying mechanisms of metabolic diseases. The peroxisome proliferator-activated receptor gamma (PPARγ) is a member of the nuclear receptor superfamily, and is vital for the transcriptional regulation of adipogenesis, insulin sensitivity and lipid metabolism. In the mouse model, it has been demonstrated that global knockout of PPARγ leads to severe metabolic disturbance, resulting in embryonic lethality. However, the specific regulatory roles of its two protein isoforms, PPARγ1 and PPARγ2, remain uncertain, due to limitations of reagents and appropriate mouse models. To investigate the hypothesis that PPARγ1 and PPARγ2 are functionally distinct, we generated PPARγ1 and PPARγ2 tagged mice using CRISPR-Cas9 technology. PPARγ1 and PPARγ2 specific knockout mice were also generated incidentally during this process, via aberrant recombination. By reverse-transcription quantitative PCR (RT-qPCR), and western blot, we confirmed the presence of the appropriate tags in our PPARγ1 and PPARγ2 tagged mice, with no significant disruption to mRNA or protein expression. Furthermore, we found that PPARγ1 mRNA and protein expression levels were reduced in our PPARγ1 knockout model, compared to the wild type. Interestingly, we found that there was a complete loss of PPARγ2 protein expression, despite an increase in PPARγ2 mRNA expression in our PPARγ2 knockout model. These data suggest that we have successfully generated PPARγ1 and PPARγ2 knockin and knockout mice. Our mouse models provide a valuable tool to study the individual roles of PPARγ1 and PPARγ2 in adipogenesis, insulin sensitivity and metabolic disease.
Comments
Presented at National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Medical Student Research Symposium, 2018