KillerRed as a Tool to Study the Cellular Responses to Peroxisome-Derived Oxidative Stress

Many biological processes and cell fate decisions are modulated by changes in redox environment. To gain insight into how subcellular compartmentalization of reactive oxygen species (ROS) formation contributes to (site-specific) redox signaling and oxidative stress responses, it is critical to have access to tools that allow tight spatial and temporal control of ROS production. Over the past decade, the use of genetically encoded photosensitizers has attracted growing interest of researchers because these proteins can be easily targeted to various subcellular compartments and allow for controlled release of ROS when excited by light. This chapter provides guidance and practical advice on the use of po-KR, a peroxisomal variant of the phototoxic red fluorescent protein KillerRed, to address fundamental questions about how mammalian cells cope with peroxisome-derived oxidative stress.