Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase
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 pho...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2021-08-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/67075 |
| _version_ | 1811252197099307008 |
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| author | Lloyd Davis Inja Radman Angeliki Goutou Ailish Tynan Kieran Baxter Zhiyan Xi Jack M O'Shea Jason W Chin Sebastian Greiss |
| author_facet | Lloyd Davis Inja Radman Angeliki Goutou Ailish Tynan Kieran Baxter Zhiyan Xi Jack M O'Shea Jason W Chin Sebastian Greiss |
| author_sort | Lloyd Davis |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T16:31:16Z |
| format | Article |
| id | doaj.art-9ed4db72b5494942b206e4f1f417c6b3 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:31:16Z |
| publishDate | 2021-08-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-9ed4db72b5494942b206e4f1f417c6b32022-12-22T03:25:08ZengeLife Sciences Publications LtdeLife2050-084X2021-08-011010.7554/eLife.67075Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinaseLloyd Davis0Inja Radman1Angeliki Goutou2Ailish Tynan3Kieran Baxter4Zhiyan Xi5Jack M O'Shea6https://orcid.org/0000-0002-9694-7340Jason W Chin7Sebastian Greiss8https://orcid.org/0000-0001-9130-0831Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United KingdomMedical Research Council Laboratory of Molecular Biology, Cambridge, United KingdomCentre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United KingdomCentre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United KingdomCentre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United KingdomCentre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United KingdomCentre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United KingdomMedical Research Council Laboratory of Molecular Biology, Cambridge, United KingdomCentre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United KingdomSynthetic 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.https://elifesciences.org/articles/67075genetic code expansionnon canonical amino acidsoptogeneticsCre recombinasephoto-caged amino acid |
| spellingShingle | Lloyd Davis Inja Radman Angeliki Goutou Ailish Tynan Kieran Baxter Zhiyan Xi Jack M O'Shea Jason W Chin Sebastian Greiss Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase eLife genetic code expansion non canonical amino acids optogenetics Cre recombinase photo-caged amino acid |
| title | Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase |
| title_full | Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase |
| title_fullStr | Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase |
| title_full_unstemmed | Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase |
| title_short | Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase |
| title_sort | precise optical control of gene expression in c elegans using improved genetic code expansion and cre recombinase |
| topic | genetic code expansion non canonical amino acids optogenetics Cre recombinase photo-caged amino acid |
| url | https://elifesciences.org/articles/67075 |
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