Corticotropin-releasing factor (CRF) is a neurotransmitter and hormone believed to integrate responses to stress. Evidence suggests central CRF systems are overactive in some individuals suffering from depression and anxiety disorders. CRF receptor antagonism blocks stress-induced endocrine, autonomic and behavioral effects in animal models, and studies have implicated the CRF₂ receptor (CRF₂) in anxiety-related behaviors. Greater understanding of the regulation of CRF₂ expression may facilitate understanding mechanisms underlying anxiety. The present studies are the first to characterize the transcriptional regulation of the human CRF_2(a), the predominant CRF₂ isoform in brain. Four kb of sequence immediately upstream of the first exon of CRF_2(a) represented our full-length promoter region. Sequentially smaller fragments of the CRF_2(a) promoter region were generated by PCR and cloned upstream of a luciferase reporter gene. Expression was monitored from these constructs within CHO-K1 cells and within A7R5 cells that express CRF₂. Glucocorticoid treatment decreased expression and elevating intracellular cAMP increased expression from the human CRF_2(a) promoter. The regions of the CRF_2(a) promoter that regulate the inducible expression were determined, and the functional CRE and GRE cis-regulatory elements within these regions were identified using a combination of site-directed mutagenesis and electrophoretic mobility shift assays. Given the possibility of species-specific differences in gene expression, interpretation of gene expression studies from rat and mouse model systems is difficult. Examination of expression from the human CRF_2(a) promoter will provide insight into these model systems and may translate more readily to the development of therapeutics to treat human psychiatric illness.