Reference Report

Reference
Citation	Mank, M., Reiff, D.F., Heim, N., Friedrich, M.W., Borst, A., Griesbeck, O. (2006). A FRET-based calcium biosensor with fast signal kinetics and high fluorescence change. Biophys. J. 90(5): 1790--1796. (Export to RIS)
FlyBase ID	FBrf0189879
Publication Type	Research paper
PubMed ID	16339891
PubMed Abstract	Genetically encoded calcium biosensors have become valuable tools in cell biology and neuroscience, but some aspects such as signal strength and response kinetics still need improvement. Here we report the generation of a FRET-based calcium biosensor employing troponin C as calcium-binding moiety that is fast, is stable in imaging experiments, and shows a significantly enhanced fluorescence change. These improvements were achieved by engineering magnesium and calcium-binding properties within the C-terminal lobe of troponin C and by the incorporation of circularly permuted variants of the green fluorescent protein. This sensor named TN-XL shows a maximum fractional fluorescence change of 400% in its emission ratio and linear response properties over an expanded calcium regime. When imaged in vivo at presynaptic motoneuron terminals of transgenic fruit flies, TN-XL exhibits highly reproducible fluorescence signals with the fastest rise and decay times of all calcium biosensors known so far.
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Language of Publication	English
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Publication Type	Journal
Abbreviation	Biophys. J.
Title	Biophysical Journal
Publication Year	1960-
ISBN/ISSN	0006-3495
Data from Reference
Alleles (8)
	Avic\GFP^{Cameleon2.0.Scer\UAS} Avic\GFP^{Cameleon3.3.Scer\UAS} Avic\GFP^{GCaMP1.3.Scer\UAS.cRa} Avic\GFP^{GCaMP1.6.Scer\UAS} Avic\GFP^{TN-L15.Scer\UAS.T:Ggal\TNNC2} Avic\GFP^{TN-XL.Scer\UAS.T:Ggal\TNNC2} Scer\GAL4^elav-C155 T:Avic\GFP^EGFP
Constructs (6)
	P{UAS-Cameleon.2.0} P{UAS-Cameleon.3.3} P{UAS-GCaMP1.3.R} P{UAS-G-CaMP.1.6} P{UAS-TN-L15} P{UAS-TN-XL}
Genes (3)
	Avic\GFP Scer\GAL4 T:Ggal\TNNC2
Insertions (1)
	P{GawB}elav^C155
Natural transposons (1)
	P-element