Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase.

Authors
Davis, Lloyd 
Radman, Inja 
Goutou, Angeliki 
Tynan, Ailish 
Baxter, Kieran 

Loading...
Thumbnail Image
Type
Article
Change log
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.

Publication Date
2021-08-05
Online Publication Date
2021-08-05
Acceptance Date
2021-08-04
Keywords
C. elegans, Cre recombinase, biochemistry, chemical biology, genetic code expansion, neuroscience, non canonical amino acids, optogenetics, photo-caged amino acid, Animals, Behavior, Animal, Caenorhabditis elegans, Gene Expression, Genetic Code, Genetic Engineering, Integrases, Neurons, Optogenetics, Touch Perception
Journal Title
Elife
Journal ISSN
2050-084X
2050-084X
Volume Title
10
Publisher
eLife Sciences Publications, Ltd