Introduction: The mitochondrial uniporter (MCU) Ca²⁺ ion channel represents the primary means for Ca²⁺ uptake by mitochondria. Mitochondrial matrix Ca²⁺ plays critical roles in mitochondrial bioenergetics by impinging upon respiration, energy production, and flux of biochemical intermediates through the TCA cycle. Inhibition of MCU in oncogenic cell lines results in an energetic crisis and reduced cell proliferation unless media is supplemented with nucleosides, pyruvate or α-KG. Nevertheless, the roles of MCU-mediated Ca²⁺ influx in cancer cells remain unclear, in part because of a lack of genetic models.