Affiliation:
1. Centre for Discovery Brain Sciences, University of Edinburgh
2. Medical Research Council Laboratory of Molecular Biology
Abstract
Synthetic strategies for optically controlling gene expression may enable the precise spatiotemporal control of genes in any combination of cells that cannot be targeted with specific promoters. We develop an improved genetic code expansion system in Caenorhabditis elegans and use it to create a photoactivatable Cre recombinase. We laser-activate Cre in single neurons within a bilaterally symmetric pair to selectively switch on expression of a loxP-controlled optogenetic channel in the targeted neuron. We use the system to dissect, in freely moving animals, the individual contributions of the mechanosensory neurons PLML/PLMR to the C. elegans touch response circuit, revealing distinct and synergistic roles for these neurons. We thus demonstrate how genetic code expansion and optical targeting can be combined to break the symmetry of neuron pairs and dissect behavioural outputs of individual neurons that cannot be genetically targeted.
Funder
H2020 European Research Council
Medical Research Council
Wellcome-Trust University of Edinburgh Institutional Strategic Support Fund ISS2
The Royal Society
Muir Maxwell Epilepsy Centre
Louis Jeantet Foundation
Herchel Smith Foundation
University of Edinburgh, Edinburgh Global Award and Principal's Career Development PhD studentship
Publisher
eLife Sciences Publications, Ltd
Subject
General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience
Cited by
10 articles.
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