Given that ribokinase (RBKS) catalyzes the key phosphorylation of D-ribose to ribose-5-phosphate, we hypothesized that the Rbks gene plays a critical role in regulating systemic glucose and lipid metabolism. To test this hypothesis, we generated Rbks-knockout (Rbks-KO) C57BL/6J mice using CRISPR/Cas9 technology and performed proteomic analysis on their livers. A total of 117 differentially expressed proteins were identified, which are primarily involved in glycolipid metabolism and oxidative phosphorylation.