Academic Journal
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 |
|---|---|
| المؤلفون: | Ast, Cindy, Foret, Jessica, Oltrogge, Luke M., De Michele, Roberto, Kleist, Thomas J., Ho, Cheng-Hsun, Frommer, Wolf B. |
| المصدر: | Nature Communications ; volume 8, issue 1 ; ISSN 2041-1723 |
| بيانات النشر: | Springer Science and Business Media LLC |
| سنة النشر: | 2017 |
| 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. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1038/s41467-017-00400-2 |
| الاتاحة: | http://dx.doi.org/10.1038/s41467-017-00400-2 https://www.nature.com/articles/s41467-017-00400-2.pdf https://www.nature.com/articles/s41467-017-00400-2 |
| Rights: | https://creativecommons.org/licenses/by/4.0 ; https://creativecommons.org/licenses/by/4.0 |
| رقم الانضمام: | edsbas.23D94F32 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1038/s41467-017-00400-2 |
|---|