Development of highly sensitive fluorescent probes for the detection of β-galactosidase activity - application to the real-time monitoring of senescence in live cells

We report the development of four novel fluorescent probes to monitor the activity of the β-galactosidase enzyme (β-gal), in vitro and in living cells. The fluorophores are based on a 6-amino-styryl-benzothiazole push-pull core and display a strong ICT emission. The probes encompass the fluorescent...

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Bibliographic Details
Published in:Analyst (London) 2018-05, Vol.143 (11), p.268-2688
Main Authors: Safir Filho, Mauro, Dao, Pascal, Gesson, Maéva, Martin, Anthony R, Benhida, Rachid
Format: Article
Language:English
Subjects:
beta-Galactosidase - metabolism
Biomarkers
Cell Line, Tumor
Cellular Senescence
Chemical compounds
Chemical Sciences
Flow Cytometry
Fluorescent Dyes
Fluorescent indicators
Galactosidase
Humans
Microscopy, Fluorescence
Real time
Senescence
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Summary:We report the development of four novel fluorescent probes to monitor the activity of the β-galactosidase enzyme (β-gal), in vitro and in living cells. The fluorophores are based on a 6-amino-styryl-benzothiazole push-pull core and display a strong ICT emission. The probes encompass the fluorescent motif that is connected to a β- d -galactopyranoside moiety through a self-immolative benzyl carbamate linker ( βGal-1-4 ). The screening of four different fluorophores enabled us to access new light-up and two-band ratiometric reporters. The four probes, βGal-1-4 , exhibited an extremely fast response and over 200-fold fluorescence enhancement ( βGal-1 ) following the enzymatic cleavage of the β- d -galactopyranoside unit. This rapid and extremely sensitive response allowed the detection of senescence-associated β-galactosidase (SA-β-gal) activity; a widely used biomarker of senescence. More importantly, βGal-1 also enabled us to monitor, in real-time, the emergence of senescence in live cells , i.e. the phenotypic transformation from normal to senescent cell. These findings underpin the fact that βGal-1 may find useful applications in biomedical research. Importantly, βGal-1 is suitable for epifluorescence and confocal microscopies, and flow cytometry techniques, which are among the most common analytical tools in biology. Highly sensitive fluorescent reporters for the detection and monitoring of β-galactosidase activity in live cells.
ISSN:0003-2654
1364-5528
DOI:10.1039/c8an00516h