Our recent genomic studies identified a complex kidney specific enhancer module located within the introns of adjacent Mettl1 (M1) and Mettl21b (M21) genes that mediate basal and PTH induction of Cyp27b1, as well as suppression by FGF23 and 1,25 dihydroxyvitamin D₃ [1,25(OH)₂D₃]. The tissue specificity for this regulatory module appears to be localized exclusively to renal proximal tubules. Gross deletion of these segments in mice has severe consequences on skeletal health, and directly affects Cyp27b1 expression in the kidney. Deletion of both the M1 and M21 submodules together almost completely eliminates basal Cyp27b1 expression in the kidney, creating a renal specific pseudo null mouse, resulting in a systemic and skeletal phenotype similar to that of the Cyp27b1 KO mouse caused by high levels of both 25 hydroxyvitamin D₃ [25(OH)D₃] and PTH and depletion of 1,25(OH)₂D₃. Cyp24a1 levels in the double KO mouse also decrease because of compensatory downregulation of the gene by elevated PTH and reduced FGF23 that is mediated by an intergenic module located downstream of the Cyp24a1 gene. Outside of the kidney in nonrenal target cells (NRTCs), expression of Cyp27b1 in these mutant mice was unaffected. Dietary normalization of calcium, phosphate, PTH, and FGF23 rescues the aberrant phenotype of this mouse and normalizes the skeleton. In addition, both the high levels of 25(OH)D₃ were reduced and the low levels of 1,25(OH)₂D₃ were fully eliminated in these mutant mice as a result of the rescue induced normalization of renal Cyp24a1. Thus, these hormone regulated enhancers for both Cyp27b1 and Cyp24a1 in the kidney are responsible for the circulating levels of 1,25(OH)₂D₃ in the blood. The retention of Cyp27b1 and Cyp24a1 expression in NRTCs of these endocrine 1,25(OH)₂D₃ deficient mice suggests that this Cyp27b1 pseudo null mouse will provide a model for the future exploration of the role of NRTC produced 1,25(OH)₂D₃ in the hormone's diverse noncalcemic actions in both health and disease. © 2020 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.