Objective In the clinical studies, mitochondrial dysfunction of skeletal muscle has been suggested as an etiological factor of insulin resistance and it is considered as a primary factor in the development of diabetes. However, the metabolic phenotypes of altered OXPHOS function in skeletal muscle remains to be identified. To investigate the skeletal muscle specific roles of OXPHOS defect, we developed mice with Crif1deficiency and analyzed the skeletal muscle and metabolic phenotypes. Methods we generated skeletal muscle-specific Crif1 knockout (Crif1 SKO) mice, crossing the Crif1f/f mice to myosin light chain (MLC)-Cre mice. To identify mitochondrial dysfunction in Crif1 SKO and WT mice, we analyzed OXPHOS gene expression, mitochondrial activity and mitochondria morphology. To analyze metabolic phenotypes, evaluated biochemical, histological and molecular markers. Results Muscle-specific Crif1 knockout (Crif1 SKO) pups were born healthy and viable, but adult mice showed progressive loss of body weight associated with reduced skeletal muscle and fat mass. Furthermore, increased mortality and decreased physical performance were observed in homozygote mice. Ultra-structural examination showed increased abnormal mitochondria and decreased OXPHOS complexes in skeletal muscle. Interestingly, these mice had improved glucose disposal rate compared to control mice. We report here that OXPHOS defect of skeletal muscle induced muscle atrophy and enhance glucose disposal. Conclusion We suggest that Crif1 is a critical factor for the biogenesis of the OXPHOS complexes and loss of Crif1 function results in dystrophic changes in skeletal muscle. But homozygotic loss of Crif1 in mice showed improvement in systemic glucose tolerance.