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<p>C57BL/6J mice are widely used in biomedical research and are susceptible to insulin resistance and dyslipidemia when challenged with high fat diets relative to other inbred strains. Interestingly, C57Bl/6J mice contain a naturally occurring premature termination codon in the lipid flippase <i>Atp10D</i>, and previous studies have linked <i>Atp10D</i> to the metabolic disease-prone phenotype in mice and atherosclerotic severity in humans. In this study, we used CRISPR/Cas9 to revert the premature termination codon to the wild-type glutamine codon (<i>Atp10D *817Q</i>) in the C57Bl/6J mouse strain. The RNA transcripts from original and corrected alleles are dually expressed in heterozygous mice, suggesting that the mutant transcript escapes nonsense-mediated decay. Expression of two corrected <i>Atp10D</i> alleles restores wildtype expression levels of the transcript and protein in the liver. When challenged with a high fat diet, <i>Atp10D</i><sup><i>−/−</i></sup> (original) and <i>Atp10D</i><sup><i>+/+</i></sup> (corrected) C57Bl/6J mice showed no significant difference in weight gain, glucose tolerance, or plasma levels of triglycerides, cholesterol, or free fatty acids. However, the female <i>Atp10D</i><sup><i>+/+</i></sup> mice displayed an increase in complex glycosphingolipids and a reduction in cardiolipins in the plasma. These results suggest that restoring the expression of <i>Atp10D</i> in C57Bl/6J mice does not reverse insulin resistance and dyslipidemia in response to high fat diet feeding.</p>

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Repairing Atp10D in C57Bl/6J mice restores protein expression but does not mitigate metabolic stress from high fat diet

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月7日

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  • Alexander J. Mansueto,
  • Adriana C. Norris,
  • Eugenia M. Yazlovitskaya,
  • Lin Zhu,
  • Joey Lu,
  • David C. Koomen,
  • Jody C. May,
  • Minsoo Kim,
  • Jake N. Hermanson,
  • Robert S. Molday,
  • John A. McLean,
  • Lars Plate,
  • John M. Stafford,
  • Todd R. Graham

摘要

C57BL/6J mice are widely used in biomedical research and are susceptible to insulin resistance and dyslipidemia when challenged with high fat diets relative to other inbred strains. Interestingly, C57Bl/6J mice contain a naturally occurring premature termination codon in the lipid flippase Atp10D, and previous studies have linked Atp10D to the metabolic disease-prone phenotype in mice and atherosclerotic severity in humans. In this study, we used CRISPR/Cas9 to revert the premature termination codon to the wild-type glutamine codon (Atp10D *817Q) in the C57Bl/6J mouse strain. The RNA transcripts from original and corrected alleles are dually expressed in heterozygous mice, suggesting that the mutant transcript escapes nonsense-mediated decay. Expression of two corrected Atp10D alleles restores wildtype expression levels of the transcript and protein in the liver. When challenged with a high fat diet, Atp10D−/− (original) and Atp10D+/+ (corrected) C57Bl/6J mice showed no significant difference in weight gain, glucose tolerance, or plasma levels of triglycerides, cholesterol, or free fatty acids. However, the female Atp10D+/+ mice displayed an increase in complex glycosphingolipids and a reduction in cardiolipins in the plasma. These results suggest that restoring the expression of Atp10D in C57Bl/6J mice does not reverse insulin resistance and dyslipidemia in response to high fat diet feeding.