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

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Abstract

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.

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

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Publication details

The article was received on 20 Mar 2018, accepted on 13 May 2018 and first published on 14 May 2018


Article type: Paper
DOI: 10.1039/C8AN00516H
Citation: Analyst, 2018, Advance Article
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    Development of highly sensitive fluorescent probes for the detection of β-galactosidase activity – application to the real-time monitoring of senescence in live cells

    M. Safir Filho, P. Dao, M. Gesson, A. R. Martin and R. Benhida, Analyst, 2018, Advance Article , DOI: 10.1039/C8AN00516H

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