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Detection of miRNA in Live Cells by Using Templated RuII‐Catalyzed Unmasking of a Fluorophore |
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| Authors: | Dr. Kalyan K. Sadhu Prof. Nicolas Winssinger |
| Affiliation: | 1. Institut de Science et Ingénierie Supramoléculaires (ISIS‐UMR 7006), Université de Strasbourg, CNRS, 8 allée Gaspard Monge, 67000 Strasbourg (France);2. Department of Organic Chemistry, University of Geneva, 30 quai Ernest Ansermet, 1211 Geneva 4 (Switzerland), Fax: (+41)?223793215 |
| Abstract: | 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 pM 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. |
| Keywords: | bioorthogonal chemistry imaging agents peptides RNA recognition templated reactions |