Cindy Ast, Jessica Foret, Luke M. Oltrogge, Roberto De Michele, Thomas J. Kleist, Cheng-Hsun Ho & Wolf B. Frommer (2017) Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins.Nature Communications(2017), doi:10.1038/s41467-017-00400-2
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.
Figure. Arabidopsis seedlings expressing MatryoshCaMP6s report root cytosolic calcium elevation in response to salt shock.
2018/07/16 10:30 AM
Dr. Jeng-Yuan Shiau (Dr. Lie-Fen Shyur's Lab)
Molecular insights of phytoagent deoxyelephantopin and its derivative against human triple negative breast cancer cell activities
Auditorium A134, Agricultural Technology Building, Academia Sinica 2018/08/22 10:30 AM
Dr. Nathaniel Robert Street (Associate Professor, Department of Plant Physiology, Umeå University, Sweden)
Systems genetics and genomics insights into complex traits in aspen and Norway spruce
Auditorium A134, Agricultural Technology Building, Agricultural Biotechnology Research Center