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

The Analyst
Mauro Safir FilhoRachid Benhida

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...Continue Reading

References

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Citations

Sep 23, 2020·The FEBS Journal·Estela González-GualdaDaniel Muñoz-Espín
Feb 20, 2021·Analytical and Bioanalytical Chemistry·Beatriz Lozano-TorresRamón Martínez-Máñez
May 27, 2021·Organic & Biomolecular Chemistry·Abhishek SainiSonu Kumar
Jul 5, 2019·Bioconjugate Chemistry·Jianjian ZhangKanyi Pu
Nov 10, 2021·Journal of Medicinal Chemistry·Jun LiuLina Cui

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Methods Mentioned

BETA
flow cytometry
column chromatography
NMR
fluorescence imaging
FACS
acylation
fluorescence
fluorescence microscopy

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