Resonance energy transfer sensitises and monitors in situ switching of LOV2-based optogenetic actuators
| العنوان: | Resonance energy transfer sensitises and monitors in situ switching of LOV2-based optogenetic actuators |
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| المؤلفون: | Li, Li-Li, Klein, Florence M, Li Greci, Lorenzo, Popinigis, Arkadiusz, Freudenberg, Florian, Courtney, Michael J |
| المساهمون: | Li, Li-Li [0000-0002-8888-153X], Li Greci, Lorenzo [0000-0002-1909-7645], Freudenberg, Florian [0000-0003-1438-3850], Courtney, Michael J. [0000-0001-8693-3933], Apollo - University of Cambridge Repository, Courtney, Michael J [0000-0001-8693-3933] |
| المصدر: | Nature Communications Nature Communications, Vol 11, Iss 1, Pp 1-18 (2020) |
| بيانات النشر: | Nature Publishing Group UK, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Male, Phototropins, Light, MAP Kinase Signaling System, Science, Recombinant Fusion Proteins, Green Fluorescent Proteins, 96/35, 96/33, 96/10, 14, Hippocampus, Article, 96/95, Fluorescence Resonance Energy Transfer, Animals, Humans, 631/61/338/469, 631/1647/2253, lcsh:Science, Synthetic biology, Neurons, Molecular Imaging, Rats, Optogenetics, 631/92/552, HEK293 Cells, Energy Transfer, 14/63, 13/95, lcsh:Q, Female, Protein design |
| الوصف: | Funder: Magnus Ehrnroothin Säätiö (Magnus Ehrnrooth Foundation); doi: https://doi.org/10.13039/501100004155 Funder: 7th Framework Programme Initial Training Networks FP7-PEOPLE-2013-ITN Project Number 608346 Project ‘Brain Imaging Return To Health’ r’Birth Funder: U.S. Department of Health & Human Services | NIH | National Cancer Institute (NCI) Funder: EC | Erasmus+; doi: https://doi.org/10.13039/501100010790 Engineered light-dependent switches provide uniquely powerful opportunities to investigate and control cell regulatory mechanisms. Existing tools offer high spatiotemporal resolution, reversibility and repeatability. Cellular optogenetics applications remain limited with diffusible targets as the response of the actuator is difficult to independently validate. Blue light levels commonly needed for actuation can be cytotoxic, precluding long-term experiments. We describe a simple approach overcoming these obstacles. Resonance energy transfer can be used to constitutively or dynamically modulate actuation sensitivity. This simultaneously offers on-line monitoring of light-dependent switching and precise quantification of activation-relaxation properties in intact living cells. Applying this approach to different LOV2-based switches reveals that flanking sequences can lead to relaxation times up to 11-fold faster than anticipated. In situ–measured parameter values guide the design of target-inhibiting actuation trains with minimal blue-light exposure, and context-based optimisation can increase sensitivity and experimental throughput a further 10-fold without loss of temporal precision. |
| وصف الملف: | application/pdf; application/zip; text/xml |
| اللغة: | English |
| تدمد: | 2041-1723 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bd0155e8e4bca6783b623e74686a46be http://europepmc.org/articles/PMC7547724 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....bd0155e8e4bca6783b623e74686a46be |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20411723 |
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