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A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection

Fluorogenic hybridization probes allow the detection of RNA and DNA sequences in homogeneous solution. Typically, one target molecule activates the fluorescence of a single probe molecule. This limits the sensitivity of nucleic acid detection. Herein, we report a self‐immolative molecular beacon (iM...

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Detalles Bibliográficos
Autores principales: Roth, Magdalena, Seitz, Oliver
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8597011/
https://www.ncbi.nlm.nih.gov/pubmed/34516006
http://dx.doi.org/10.1002/chem.202102600
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author Roth, Magdalena
Seitz, Oliver
author_facet Roth, Magdalena
Seitz, Oliver
author_sort Roth, Magdalena
collection PubMed
description Fluorogenic hybridization probes allow the detection of RNA and DNA sequences in homogeneous solution. Typically, one target molecule activates the fluorescence of a single probe molecule. This limits the sensitivity of nucleic acid detection. Herein, we report a self‐immolative molecular beacon (iMB) that escapes the one‐target/one‐probe paradigm. The iMB probe includes a photoreductively cleavable N‐alkyl‐picolinium (NAP) linkage within the loop region. A fluorophore at the 5’‐end serves, on the one hand, as a reporter group and, on the other hand, as a photosensitizer of a NAP‐linker cleavage reaction. In the absence of target, the iMB adopts a hairpin shape. Quencher groups prevent photo‐induced cleavage. The iMB opens upon hybridization with a target, and both fluorescent emission as well as photo‐reductive cleavage of the NAP linker can occur. In contrast to previous chemical amplification reactions, iMBs are unimolecular probes that undergo cleavage leading to products that have lower target affinity than the probes before reaction. Aided by catalysis, the method allowed the detection of 5 pm RNA target within 100 min.
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spelling pubmed-85970112021-11-22 A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection Roth, Magdalena Seitz, Oliver Chemistry Communications Fluorogenic hybridization probes allow the detection of RNA and DNA sequences in homogeneous solution. Typically, one target molecule activates the fluorescence of a single probe molecule. This limits the sensitivity of nucleic acid detection. Herein, we report a self‐immolative molecular beacon (iMB) that escapes the one‐target/one‐probe paradigm. The iMB probe includes a photoreductively cleavable N‐alkyl‐picolinium (NAP) linkage within the loop region. A fluorophore at the 5’‐end serves, on the one hand, as a reporter group and, on the other hand, as a photosensitizer of a NAP‐linker cleavage reaction. In the absence of target, the iMB adopts a hairpin shape. Quencher groups prevent photo‐induced cleavage. The iMB opens upon hybridization with a target, and both fluorescent emission as well as photo‐reductive cleavage of the NAP linker can occur. In contrast to previous chemical amplification reactions, iMBs are unimolecular probes that undergo cleavage leading to products that have lower target affinity than the probes before reaction. Aided by catalysis, the method allowed the detection of 5 pm RNA target within 100 min. John Wiley and Sons Inc. 2021-09-13 2021-10-13 /pmc/articles/PMC8597011/ /pubmed/34516006 http://dx.doi.org/10.1002/chem.202102600 Text en © 2021 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Communications
Roth, Magdalena
Seitz, Oliver
A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection
title A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection
title_full A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection
title_fullStr A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection
title_full_unstemmed A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection
title_short A Self‐immolative Molecular Beacon for Amplified Nucleic Acid Detection
title_sort self‐immolative molecular beacon for amplified nucleic acid detection
topic Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8597011/
https://www.ncbi.nlm.nih.gov/pubmed/34516006
http://dx.doi.org/10.1002/chem.202102600
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