Abstract
Reactive oxygen species (ROS) can modify and damage DNA. Here we report an optogenetic mutagenesis approach that is free of toxic chemicals and easy to perform by taking advantage of a genetically encoded ROS generator. This method relies on the potency of ROS generation by His-mSOG, the mini singlet oxygen generator, miniSOG, fused to a histone. Caenorhabditis elegans expressing His-mSOG in the germline behave and reproduce normally, without photoinduction. Following exposure to blue light, the His-mSOG animals produce progeny with a wide range of heritable phenotypes. We show that optogenetic mutagenesis by His-mSOG induces a broad spectrum of mutations including single-nucleotide variants (SNVs), chromosomal deletions, as well as integration of extrachromosomal transgenes, which complements those derived from traditional chemical or radiation mutagenesis. The optogenetic mutagenesis expands the toolbox for forward genetic screening and also provides direct evidence that nuclear ROS can induce heritable and specific genetic mutations.
Highlights
Reactive oxygen species (ROS) can modify and damage DNA
Among the F2 progeny from the light-treated His-mSOG parents, we found mutant animals with visible phenotypes such as uncoordinated and dumpy (Fig. 1c)
Wild-type animals treated with blue light did not produce visible mutants, nor did His-mSOG animals without exposure to blue light (Fig. 1e), indicating that the observed mutagenesis is dependent on the combination of blue-light exposure and the His-mSOG transgene
Summary
We report an optogenetic mutagenesis approach that is free of toxic chemicals and easy to perform by taking advantage of a genetically encoded ROS generator. This method relies on the potency of ROS generation by His-mSOG, the mini singlet oxygen generator, miniSOG, fused to a histone. The ability of miniSOG to locally generate ROS has been exploited to ablate cells[6] and to inactivate proteins[7] in a blue light dependent manner. We demonstrate that miniSOG fused to a histone (His-mSOG) induces heritable mutations in a light-dependent manner in C. elegans. Our observations demonstrate that high levels of ROS in the nucleus can cause specific and genetically heritable mutations
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