Mapping the Fate of Hypoxic Cells Using an Irreversible Fluorescent Switch

Methods Mol Biol. 2024:2755:49-61. doi: 10.1007/978-1-0716-3633-6_3.

Abstract

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.

Keywords: Cre-loxp; Fluorescent reporter; Hypoxia fate-mapping; Hypoxia reporter; Intratumoral hypoxia; Metastasis.

MeSH terms

  • Animals
  • Cell Line
  • Coloring Agents
  • Embryonic Development
  • Humans
  • Hypoxia*
  • Mammals
  • Mice
  • Oxygen*

Substances

  • Oxygen
  • Coloring Agents