Reference Report

Reference
Citation	Bardet, P.L., Kolahgar, G.I., Mynett, A., Miguel-Aliaga, I., Briscoe, J., Meier, P., Vincent, J.P. (2008). A fluorescent reporter of caspase activity for live imaging. Proc. Natl. Acad. Sci. U.S.A. 105(37): 13901--13905. (Export to RIS)
FlyBase ID	FBrf0205998
Publication Type	Research paper
PubMed ID	18779587
PubMed Abstract	There is a growing interest in the mechanisms that control the apoptosis cascade during development and adult life. To investigate the regulatory events that trigger apoptosis in whole tissues, we have devised a genetically encoded caspase sensor that can be detected in live and fixed tissue by standard confocal microscopy. The sensor comprises two fluorophores, mRFP, monomeric red fluorescent protein (mRFP) and enhanced green fluorescent protein (eGFP), that are linked by an efficient and specific caspase-sensitive site. Upon caspase activation, the sensor is cleaved and eGFP translocates to the nucleus, leaving mRFP at membranes. This is detected before other markers of apoptosis, including anti-cleaved caspase 3 immunoreactivity. Moreover, the sensor does not perturb normal developmental apoptosis and is specific, as cleavage does not occur in Drosophila embryos that are unable to activate the apoptotic cascade. Importantly, dying cells can be recognized in live embryos, thus opening the way for in vivo imaging. As expected from the high conservation of caspases, it is also cleaved in dying cells of chick embryos. It is therefore likely to be generally useful to track the spatiotemporal pattern of caspase activity in a variety of species.
DOI	10.1073/pnas.0806983105
Related Publication(s)
Personal communication to FlyBase	UAS-Apoliner. Vincent, 2010.9.7, UAS-Apoliner. [FBrf0211799]
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Language of Publication	English
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Parent Publication
Publication Type	Journal
Abbreviation	Proc. Natl. Acad. Sci. U.S.A.
Title	Proceedings of the National Academy of Sciences of the United States of America
Publication Year	1915-
ISBN/ISSN	0027-8424
Data from Reference
Aberrations (1)
	Df(3L)H99
Alleles (8)
	Scer\GAL4^69B Scer\GAL4^tub.PU Scer\GAL4^Vap.P0201 T:Avic\GFP^EGFP T:Disc\RFP^mRFP th^{D20A.αTub84B.T:Mmus\Cd8a.T:Disc\RFP-mRFP.T:SV40\nls2.T:Avic\GFP-EGFP} th^{N21G.N22V.Scer\UAS.T:Mmus\Cd8a.T:Disc\RFP-mRFP.T:SV40\nls2.T:Avic\GFP-EGFP} th^{N21G.N22V.αTub84B.T:Mmus\Cd8a.T:Disc\RFP-mRFP.T:SV40\nls2.T:Avic\GFP-EGFP}
Constructs (4)
	P{Tub-Apoliner} P{Tub-Apomut} P{tub-GAL4.U} P{UAS-Apoliner}
Genes (9)
	BacA\p35 decay Ice rpr Scer\GAL4 T:Mmus\Cd8a T:SV40\nls2 th αTub84B
Insertions (3)
	P{GAL4}Vap.P0201 P{GawB}69B P{UAS-Apoliner}5
Natural transposons (1)
	P-element