Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins
| العنوان: | Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins |
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| المؤلفون: | Cheng-Hsun Ho, Thomas J. Kleist, Wolf B. Frommer, Luke M. Oltrogge, Jessica Foret, Roberto De Michele, Cindy Ast |
| المصدر: | Nature Communications, Vol 8, Iss 1, Pp 1-13 (2017) Nature communications 8 (2017). doi:10.1038/s41467-017-00400-2 info:cnr-pdr/source/autori:Ast, Cindy; Foret, Jessica; Oltrogge, Luke M.; De Michele, Roberto; Kleist, Thomas J.; Ho, Cheng-Hsun; Frommer, Wolf B./titolo:Ratiometric Matryoshka biosensors from a nested cassette of green-and orange-emitting fluorescent proteins/doi:10.1038%2Fs41467-017-00400-2/rivista:Nature communications/anno:2017/pagina_da:/pagina_a:/intervallo_pagine:/volume:8 Nature Communications |
| بيانات النشر: | Nature Portfolio, 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0106 biological sciences, 0301 basic medicine, genetic structures, Science, Arabidopsis, ammonium transporter, General Physics and Astronomy, Nanotechnology, Biosensing Techniques, sensors, 01 natural sciences, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Ammonium Compounds, Humans, Fluorescent protein, lcsh:Science, Multidisciplinary, calcium, Extramural, Dynamic range, Chemistry, technology, industry, and agriculture, Biological Transport, General Chemistry, biosensors, Fluorescence, Luminescent Proteins, HEK293 Cells, 030104 developmental biology, GCaMP, lcsh:Q, fluorescence, Biosensor, 010606 plant biology & botany, Ca2 imaging, Ammonium transport |
| الوصف: | Sensitivity, dynamic and detection range as well as exclusion of expression and instrumental artifacts are critical for the quantitation of data obtained with fluorescent protein (FP)-based biosensors in vivo. Current biosensors designs are, in general, unable to simultaneously meet all these criteria. Here, we describe a generalizable platform to create dual-FP biosensors with large dynamic ranges by employing a single FP-cassette, named GO-(Green-Orange) Matryoshka. The cassette nests a stable reference FP (large Stokes shift LSSmOrange) within a reporter FP (circularly permuted green FP). GO- Matryoshka yields green and orange fluorescence upon blue excitation. As proof of concept, we converted existing, single-emission biosensors into a series of ratiometric calcium sensors (MatryoshCaMP6s) and ammonium transport activity sensors (AmTryoshka1;3). We additionally identified the internal acid-base equilibrium as a key determinant of the GCaMP dynamic range. Matryoshka technology promises flexibility in the design of a wide spectrum of ratiometric biosensors and expanded in vivo applications. Single fluorescent protein biosensors are susceptible to expression and instrumental artifacts. Here Ast et al. describe a dual fluorescent protein design whereby a reference fluorescent protein is nested within a reporter fluorescent protein to control for such artifacts while preserving sensitivity and dynamic range. |
| اللغة: | English |
| تدمد: | 2041-1723 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1beab4368e4b7cedb95b76c561c79a00 https://doaj.org/article/af5004125dba42869f7a394844689d77 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....1beab4368e4b7cedb95b76c561c79a00 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20411723 |
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