Mapping the Fate of Hypoxic Cells Using an Irreversible Fluorescent Switch.

Hypoxia has been reported to promote tumor progression and metastasis in murine models, and patients with hypoxic tumors have a worse prognosis. Besides its effect on cancer, normal processes like embryogenesis, or other pathologies such as ischemia, depend on hypoxia-regulated mechanisms. Given the degradable nature of HIF-1/2α in the presence of oxygen, defining the role of hypoxia in modeling biological processes becomes challenging when a cell enters oxygen-rich regions within a tissue. Here, we describe a unique approach to permanently mark cells that experience hypoxia with a fluorescent protein switch that is maintained even after a cell is reoxygenated. This method consists of a dual-viral delivery system that can be transduced into any mammalian cell line.