Highly efficient optogenetic cell ablation in C. elegans using membrane-targeted miniSOG
authors: Suhong Xu, Andrew D. Chisholm
doi: 10.1038/srep21271
CITATION
Xu, S., & Chisholm, A. D. (2016). Highly efficient optogenetic cell ablation in C. elegans using membrane-targeted miniSOG. Scientific Reports, 6(1), 21271. https://doi.org/10.1038/srep21271
ABSTRACT
The genetically encoded photosensitizer miniSOG (mini Singlet Oxygen Generator) can be used to kill cells in C. elegans. miniSOG generates the reactive oxygen species (ROS) singlet oxygen after illumination with blue light. Illumination of neurons expressing miniSOG targeted to the outer mitochondrial membrane (mito-miniSOG) causes neuronal death. To enhance miniSOG’s efficiency as an ablation tool in multiple cell types we tested alternative targeting signals. We find that membrane targeted miniSOG allows highly efficient cell killing. When combined with a point mutation that increases miniSOG’s ROS generation, membrane targeted miniSOG can ablate neurons in less than one tenth the time of mito-miniSOG. We extend the miniSOG ablation technique to non-neuronal tissues, revealing an essential role for the epidermis in locomotion. These improvements expand the utility and throughput of optogenetic cell ablation in C. elegans.
fleeting notes
miniSOG
- used to kill cells in c elegans
PH-miniSOG optogenetically ablates cells
Methods used:
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2Hz illumination was most effective (.25s on .25s off)
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~2mW/mm2
highlights
“After 1 min continuous blue light illumination, 80% of Pcol-19-PH-miniSOG transgenic animals were paralyzed”Page 2
“found 2 Hz (0.25 sec on and 0.25 sec off) to be most effective in paralyzing epidermal PH-miniSOG transgenic animals (Figure S1e). Unless stated, in subsequent experiments we use 2 Hz illumination.”Page 2
“2 min blue light was required to paralyze 50% of animals expressing PH-miniSOG at comparable levels”Page 2
“2 h after illumination, 100% of PH-miniSOG animals were paralyzed”Page 2
“and eventually died (defined as nonresponsive to touch) within the next 4–6 h”Page 2
“hereas PH-miniSOG has stronger acute effects.”Page 2
“2.01 mW/mm2”Page 11