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Molecular magnetic resonance imaging (MRI) is an emerging field that is set to revolutionize our perspective of disease diagnosis, treatment efficacy monitoring, and precision medicine in full concordance with personalized medicine. A wide range of hyperpolarized (HP) ¹²⁹Xe biosensors have been recently developed, demonstrating their potential applications in molecular settings, and achieving notable success within <i>in vitro</i> studies. The favorable nuclear magnetic resonance properties of ¹²⁹Xe, coupled with its non-toxic nature, high solubility in biological tissues, and capacity to dissolve in blood and diffuse across membranes, highlight its superior role for applications in molecular MRI settings. The incorporation of reporters that combine signal enhancement from both hyperpolarized ¹²⁹Xe and chemical exchange saturation transfer holds the potential to address the primary limitation of low sensitivity observed in conventional MRI. This review provides a summary of the various applications of HP ¹²⁹Xe biosensors developed over the last decade, specifically highlighting their use in MRI. Moreover, this paper addresses the evolution of <i>in vivo</i> applications of HP ¹²⁹Xe, discussing its potential transition into clinical settings.