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Detection of miRNA in Live Cells by Using Templated Ru II ‐Catalyzed Unmasking of a Fluorophore
Author(s) -
Sadhu Kalyan K.,
Winssinger Nicolas
Publication year - 2013
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201300060
Subject(s) - fluorophore , nucleic acid , linker , hela , chemistry , rhodamine , cleavage (geology) , transfection , fluorescence , biophysics , combinatorial chemistry , biochemistry , biology , cell , gene , paleontology , physics , quantum mechanics , fracture (geology) , computer science , operating system
Reactions templated by cellular nucleic acids are attractive for nucleic acid sensing or responsive systems. Herein we report the use of a photocatalyzed reductive cleavage of an immolative linker to unmask a rhodamine fluorophore, and its application to miRNA imaging. The reaction was found to proceed with a very high turnover (>4000) and provided reliable detection down to 5 p M of template by using γ‐serine‐modified peptide nucleic acid (PNA) probes. The reaction was used for the selective detection of miR‐21 in BT474 cells and miR‐31 in HeLa cells following irradiation for 30 min. The probes were introduced by using reversible permeation with streptolysin‐O (SLO) or a transfection technique.