Hepcidin (HAMP) is synthesized in the liver and characterizes the key iron-regulatory hormone that controls systemic iron homeostasis. Cereblon (CRBN) and Kruppel-like factor 15 (KLF15) are known to regulate diverse physiological functions. In this study, we investigate the role of CRBN on hepatic hepcidin gene expression and its production under gluconeogenic stimuli. Fasted mice as well as forskolin (FSK)- and glucagon (GLU)-treated mice had reduced serum iron levels and increased expressions of hepatic Crbn and Klf15 and hepcidin secretion. MicroRNA (miRNA) expression analysis in fasted and Ad-Crbn-infected mice exhibited significant reduction in microRNA-639 (miR-639). Hepatic overexpression of Crbn elevated hepcidin expression and its production along with Klf15 gene expression, whereas knockdown of Crbn and Klf15 markedly decreased the FSK- and fasting-mediated induction of hepcidin gene expression and its biosynthesis in mouse livers and primary hepatocytes. Moreover, expression of KLF15 significantly increased the hepcidin reporter gene activity, and it was exclusively dependent on the KLF15-binding site identified within the hepcidin gene promoter. Overall, this study demonstrates that CRBN and KLF15 are novel mediators of gluconeogenic signal-induced hepcidin gene expression and its production. CRBN and KLF15 may be novel potential therapeutic targets to intervene the metabolic dysfunction.