Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions
The kinetics of a ligand-receptor interaction determine the responses of the receptor-expressing cell. One approach to experimentally and reversibly change this kinetics on demand is optogenetics. We have previously developed a system in which the interaction of a modified receptor with an engineere...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-02-01
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| Series: | Frontiers in Molecular Biosciences |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2023.1143274/full |
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| author | Marissa Russ Marissa Russ Anna K. Ehret Anna K. Ehret Anna K. Ehret Maximilian Hörner Daniel Peschkov Rebecca Bohnert Vincent Idstein Vincent Idstein Vincent Idstein Susana Minguet Susana Minguet Wilfried Weber Björn F. Lillemeier O. Sascha Yousefi Wolfgang W. Schamel Wolfgang W. Schamel |
| author_facet | Marissa Russ Marissa Russ Anna K. Ehret Anna K. Ehret Anna K. Ehret Maximilian Hörner Daniel Peschkov Rebecca Bohnert Vincent Idstein Vincent Idstein Vincent Idstein Susana Minguet Susana Minguet Wilfried Weber Björn F. Lillemeier O. Sascha Yousefi Wolfgang W. Schamel Wolfgang W. Schamel |
| author_sort | Marissa Russ |
| collection | DOAJ |
| description | The kinetics of a ligand-receptor interaction determine the responses of the receptor-expressing cell. One approach to experimentally and reversibly change this kinetics on demand is optogenetics. We have previously developed a system in which the interaction of a modified receptor with an engineered ligand can be controlled by light. In this system the ligand is a soluble Phytochrome B (PhyB) tetramer and the receptor is fused to a mutated PhyB-interacting factor (PIFS). However, often the natural ligand is not soluble, but expressed as a membrane protein on another cell. This allows ligand-receptor interactions in two dimensions. Here, we developed a strategy to generate cells that display PhyB as a membrane-bound protein by expressing the SpyCatcher fused to a transmembrane domain in HEK-293T cells and covalently coupling purified PhyB-SpyTag to these cells. As proof-of-principle, we use Jurkat T cells that express a GFP-PIFS-T cell receptor and show that these cells can be stimulated by the PhyB-coupled HEK-293T cells in a light dependent manner. Thus, we call the PhyB-coupled cells opto-antigen presenting cells (opto-APCs). Our work expands the toolbox of optogenetic technologies, allowing two-dimensional ligand-receptor interactions to be controlled by light. |
| first_indexed | 2024-04-10T06:28:55Z |
| format | Article |
| id | doaj.art-ceac173bb5db4dc798bfa3529829a5dc |
| institution | Directory Open Access Journal |
| issn | 2296-889X |
| language | English |
| last_indexed | 2024-04-10T06:28:55Z |
| publishDate | 2023-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-ceac173bb5db4dc798bfa3529829a5dc2023-03-01T15:12:25ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2023-02-011010.3389/fmolb.2023.11432741143274Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactionsMarissa Russ0Marissa Russ1Anna K. Ehret2Anna K. Ehret3Anna K. Ehret4Maximilian Hörner5Daniel Peschkov6Rebecca Bohnert7Vincent Idstein8Vincent Idstein9Vincent Idstein10Susana Minguet11Susana Minguet12Wilfried Weber13Björn F. Lillemeier14O. Sascha Yousefi15Wolfgang W. Schamel16Wolfgang W. Schamel17Signalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Centre Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Centre Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, GermanySpemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Centre Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, GermanySpemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Centre Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanySignalling Research Centres BIOSS and CIBSS, Faculty of Biology, University of Freiburg, Freiburg, GermanyCentre for Chronic Immunodeficiency (CCI), Medical Centre Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, GermanyThe kinetics of a ligand-receptor interaction determine the responses of the receptor-expressing cell. One approach to experimentally and reversibly change this kinetics on demand is optogenetics. We have previously developed a system in which the interaction of a modified receptor with an engineered ligand can be controlled by light. In this system the ligand is a soluble Phytochrome B (PhyB) tetramer and the receptor is fused to a mutated PhyB-interacting factor (PIFS). However, often the natural ligand is not soluble, but expressed as a membrane protein on another cell. This allows ligand-receptor interactions in two dimensions. Here, we developed a strategy to generate cells that display PhyB as a membrane-bound protein by expressing the SpyCatcher fused to a transmembrane domain in HEK-293T cells and covalently coupling purified PhyB-SpyTag to these cells. As proof-of-principle, we use Jurkat T cells that express a GFP-PIFS-T cell receptor and show that these cells can be stimulated by the PhyB-coupled HEK-293T cells in a light dependent manner. Thus, we call the PhyB-coupled cells opto-antigen presenting cells (opto-APCs). Our work expands the toolbox of optogenetic technologies, allowing two-dimensional ligand-receptor interactions to be controlled by light.https://www.frontiersin.org/articles/10.3389/fmolb.2023.1143274/fullT cell receptorligandreceptoroptogeneticsinteractionphytochrome B |
| spellingShingle | Marissa Russ Marissa Russ Anna K. Ehret Anna K. Ehret Anna K. Ehret Maximilian Hörner Daniel Peschkov Rebecca Bohnert Vincent Idstein Vincent Idstein Vincent Idstein Susana Minguet Susana Minguet Wilfried Weber Björn F. Lillemeier O. Sascha Yousefi Wolfgang W. Schamel Wolfgang W. Schamel Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions Frontiers in Molecular Biosciences T cell receptor ligand receptor optogenetics interaction phytochrome B |
| title | Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions |
| title_full | Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions |
| title_fullStr | Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions |
| title_full_unstemmed | Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions |
| title_short | Opto-APC: Engineering of cells that display phytochrome B on their surface for optogenetic studies of cell-cell interactions |
| title_sort | opto apc engineering of cells that display phytochrome b on their surface for optogenetic studies of cell cell interactions |
| topic | T cell receptor ligand receptor optogenetics interaction phytochrome B |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2023.1143274/full |
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