Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells
Light-inducible dimerization protein modules enable precise temporal and spatial control of biological processes in non-invasive fashion. Among them, Magnets are small modules engineered from the Neurospora crassa photoreceptor Vivid by orthogonalizing the homodimerization interface into complementa...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2020-11-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/63230 |
| _version_ | 1818028539038924800 |
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| author | Lorena Benedetti Jonathan S Marvin Hanieh Falahati Andres Guillén-Samander Loren L Looger Pietro De Camilli |
| author_facet | Lorena Benedetti Jonathan S Marvin Hanieh Falahati Andres Guillén-Samander Loren L Looger Pietro De Camilli |
| author_sort | Lorena Benedetti |
| collection | DOAJ |
| description | Light-inducible dimerization protein modules enable precise temporal and spatial control of biological processes in non-invasive fashion. Among them, Magnets are small modules engineered from the Neurospora crassa photoreceptor Vivid by orthogonalizing the homodimerization interface into complementary heterodimers. Both Magnets components, which are well-tolerated as protein fusion partners, are photoreceptors requiring simultaneous photoactivation to interact, enabling high spatiotemporal confinement of dimerization with a single excitation wavelength. However, Magnets require concatemerization for efficient responses and cell preincubation at 28°C to be functional. Here we overcome these limitations by engineering an optimized Magnets pair requiring neither concatemerization nor low temperature preincubation. We validated these ‘enhanced’ Magnets (eMags) by using them to rapidly and reversibly recruit proteins to subcellular organelles, to induce organelle contacts, and to reconstitute OSBP-VAP ER-Golgi tethering implicated in phosphatidylinositol-4-phosphate transport and metabolism. eMags represent a very effective tool to optogenetically manipulate physiological processes over whole cells or in small subcellular volumes. |
| first_indexed | 2024-12-10T05:05:24Z |
| format | Article |
| id | doaj.art-b309da32d02f469c8c76ebb8197e3475 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T05:05:24Z |
| publishDate | 2020-11-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-b309da32d02f469c8c76ebb8197e34752022-12-22T02:01:16ZengeLife Sciences Publications LtdeLife2050-084X2020-11-01910.7554/eLife.63230Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cellsLorena Benedetti0https://orcid.org/0000-0003-3510-0258Jonathan S Marvin1Hanieh Falahati2Andres Guillén-Samander3Loren L Looger4https://orcid.org/0000-0002-7531-1757Pietro De Camilli5https://orcid.org/0000-0001-9045-0723Department of Neuroscience and Cell Biology, Yale University School of Medicine, New Haven, United States; Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United StatesHoward Hughes Medical Institute, Janelia Research Campus, Ashburn, United StatesDepartment of Neuroscience and Cell Biology, Yale University School of Medicine, New Haven, United States; Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United StatesDepartment of Neuroscience and Cell Biology, Yale University School of Medicine, New Haven, United States; Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United StatesHoward Hughes Medical Institute, Janelia Research Campus, Ashburn, United StatesDepartment of Neuroscience and Cell Biology, Yale University School of Medicine, New Haven, United States; Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, United States; Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, United States; Program in Cellular Neuroscience, Neurodegeneration and Repair, Yale University School of Medicine, New Haven, United StatesLight-inducible dimerization protein modules enable precise temporal and spatial control of biological processes in non-invasive fashion. Among them, Magnets are small modules engineered from the Neurospora crassa photoreceptor Vivid by orthogonalizing the homodimerization interface into complementary heterodimers. Both Magnets components, which are well-tolerated as protein fusion partners, are photoreceptors requiring simultaneous photoactivation to interact, enabling high spatiotemporal confinement of dimerization with a single excitation wavelength. However, Magnets require concatemerization for efficient responses and cell preincubation at 28°C to be functional. Here we overcome these limitations by engineering an optimized Magnets pair requiring neither concatemerization nor low temperature preincubation. We validated these ‘enhanced’ Magnets (eMags) by using them to rapidly and reversibly recruit proteins to subcellular organelles, to induce organelle contacts, and to reconstitute OSBP-VAP ER-Golgi tethering implicated in phosphatidylinositol-4-phosphate transport and metabolism. eMags represent a very effective tool to optogenetically manipulate physiological processes over whole cells or in small subcellular volumes.https://elifesciences.org/articles/63230LOV domainlight-dependent dimerizersorganelle contactsVAPcontact sitesVivid |
| spellingShingle | Lorena Benedetti Jonathan S Marvin Hanieh Falahati Andres Guillén-Samander Loren L Looger Pietro De Camilli Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells eLife LOV domain light-dependent dimerizers organelle contacts VAP contact sites Vivid |
| title | Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells |
| title_full | Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells |
| title_fullStr | Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells |
| title_full_unstemmed | Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells |
| title_short | Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells |
| title_sort | optimized vivid derived magnets photodimerizers for subcellular optogenetics in mammalian cells |
| topic | LOV domain light-dependent dimerizers organelle contacts VAP contact sites Vivid |
| url | https://elifesciences.org/articles/63230 |
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