The correlation between pH levels and diseases such as renal and heart failure, cancer, and stroke has long been acknowledged. However, the precise manipulation of pH in specific cellular locales poses a challenge, impeding the comprehensive understanding of the underlying mechanisms. Addressing this gap, we recently introduced Chemogenetic Operation of iNTRacellular prOton Levels (pH-CONTROL) offering heightened spatio-temporal resolution for pH manipulation. We thoroughly characterized this method through extensive in vitro and in cellulo investigations utilizing the SypHer3s biosensor. pH-CONTROL exhibits remarkable precision in modulating pH within distinct subcellular compartments, including the mitochondria, nucleus, and cytosol. Preliminary Seahorse data unveiled that the application of pH-CONTROL to cancer cells, particularly those expressing it in the mitochondrial intermembrane space, causes an augmentation in ATP production, as evidenced by an increased oxygen consumption rate. These findings underscore the potential of pH-CONTROL to reshape diverse cellular functions, with a particular emphasis on energy metabolism. The ability to precisely manipulate pH at the subcellular level opens up new avenues in cellular bioenergetics research, offering insights into the intricate interplay between pH dynamics and cellular processes. With its enhanced precision and versatility, pH-CONTROL holds promise for unraveling the connections between redox biology and various health and disease states. This method thus represents a valuable tool in advancing our understanding of cellular bioenergetics in the context of redox biology, paving the way for future investigations and therapeutic interventions.