--- - |2+ Abstract Aim In acute kidney injury (AKI), regions of the kidney are hypoxic. However, for reasons yet unknown, adaptation to hypoxia through hypoxia‐inducible factor (HIF) is limited. Here, we studied miR‐22, a potential HIF repressor, in normal kidneys, as well as in rhabdomyolysis‐induced AKI, a condition where miR‐22 is up‐regulated. Methods AKI in mice was provoked by IM injection of glycerol. Tissue homogenates were processed to determine the levels of candidate RNAs and proteins, as well as global gene expression profiles. Reporter assays quantified in vitro miR‐22 activity and its modulation by mimic or inhibitor molecules, under normoxia or hypoxia (1% O2) respectively. In vivo, anti–miR‐22 molecules were applied to normal mice or prior to induction of AKI. Renal outcome was assessed by measuring plasma creatinine, plasma urea and the levels of the injury markers Kim‐1 and Ngal. Results Renal miR‐22 is inducible by hypoxia and represses hypoxia‐inducible factor (HIF). Specific inhibition of miR‐22 regulates 1913 gene transcripts in kidneys controls and 3386 in AKI, many of which are involved in development or carcinogenesis. Specific inhibition of miR‐22 up‐regulates tissue protective HIF target genes, yet renal function and injury markers are unchanged or worsened. Conclusions miR‐22 is a HIF repressor constitutively expressed in the adult kidney and up‐regulated in AKI. Specific inhibition of miR‐22 is efficient in vivo and profoundly affects renal gene expression in health and disease, including up‐regulation of HIF. However, the net effect on rhabdomyolysis‐induced AKI outcome is neutral or even negative. - Acta Physiologica, Volume 224, Issue 3, November 2018.