Another class of optogenetic tools is a group of proteins that generate reactive oxygen species (ROS) when illuminated 2. Proteins such as channelrhodopsin and halorhodopsin conduct ions in response to light and have been used to control neuronal activity 1. Optogenetics uses light-sensitive proteins to induce a cellular response. Spatio-temporally controlled ROS generation will expand our understanding of how the respiratory chain and mitochondrial ROS influence whole organism physiology.

In conclusion, miniSOG-mediated CALI is a novel genetic platform for acute inactivation of respiratory chain components. Moreover, light-inducible phenotypes in vivo demonstrated that complex II activity is important under conditions of high energy demand, and that specific cell types are uniquely susceptible to loss of complex II.

Complex II activity was inactivated by blue light in mitochondria from strains expressing active miniSOG fusions, but not those from inactive fusions. The resulting mev-1::miniSOG transgene complemented mev-1 mutant phenotypes in kn1 missense and tm1081( lf) deletion mutants. elegans, using Mos1-mediated Single Copy Insertion. As proof-of-concept, we expressed the ‘singlet oxygen generator’ miniSOG as a fusion with the SDHC subunit of respiratory complex II, encoded by mev-1 in C. Chromophore-Assisted Light Inactivation (CALI) using genetically-encoded photosensitizers provides an opportunity to determine how individual respiratory chain components contribute to physiology following acute lf. Most Caenorhabditis elegans loss-of-function ( lf) mutants in nuclear-encoded components of the respiratory chain are non-viable, emphasizing the importance of respiratory function.

Mitochondria play critical roles in meeting cellular energy demand, in cell death, and in reactive oxygen species (ROS) and stress signaling.

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