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Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide

Potassium-sensing oligonucleotide, PSO, a conjugate of a quadruplex structure-forming oligonucleotide with a peptide incorporating a Förster Resonance Energy Transfer (FRET) chromophore pair, has been developed for fluorescent detection of potassium ion (K(+)) in aqueous medium. PSO 1 could be intro...

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Autores principales: Sato, Shinobu, Ohzawa, Shinsuke, Sota, Kojiro, Sakamoto, Naoto, Udo, Ayano, Sueda, Shinji, Matsuda, Tomoki, Nagai, Takeharu, Takenaka, Shigeori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9306769/
https://www.ncbi.nlm.nih.gov/pubmed/35873036
http://dx.doi.org/10.3389/fchem.2022.922094
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author Sato, Shinobu
Ohzawa, Shinsuke
Sota, Kojiro
Sakamoto, Naoto
Udo, Ayano
Sueda, Shinji
Matsuda, Tomoki
Nagai, Takeharu
Takenaka, Shigeori
author_facet Sato, Shinobu
Ohzawa, Shinsuke
Sota, Kojiro
Sakamoto, Naoto
Udo, Ayano
Sueda, Shinji
Matsuda, Tomoki
Nagai, Takeharu
Takenaka, Shigeori
author_sort Sato, Shinobu
collection PubMed
description Potassium-sensing oligonucleotide, PSO, a conjugate of a quadruplex structure-forming oligonucleotide with a peptide incorporating a Förster Resonance Energy Transfer (FRET) chromophore pair, has been developed for fluorescent detection of potassium ion (K(+)) in aqueous medium. PSO 1 could be introduced into cells for real-time imaging of cytoplasmic K(+) concentrations. To perform fluorescent imaging of K(+) on the cell surface, we synthesized twelve PSO derivatives with different types of peptide types and lengths, and oligonucleotide sequences including thrombin-binding aptamer (TBA) sequences with FAM and TAMRA as a FRET chromophore pair, and evaluated their performance. 1 was shown to respond selectively to K(+), not to most ions present in vivo, and to show reciprocal fluorescence changes in response to K(+) concentration. For the peptide chains and oligonucleotide sequences examined in this study, the PSO derivatives had K (d) values for K(+) in the range of 5–30 mM. All PSO derivatives showed high K(+) selectivity even in the presence of excess Na(+). The PSO derivatives were successfully localized to the cell surface by biotinylated concanavalin A (ConA) or sulfo-NHS-biotin via streptavidin (StAv). Fluorescence imaging of extracellular K(+) upon addition of apoptosis inducers was successfully achieved by 1 localized to the cell surface.
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spelling pubmed-93067692022-07-23 Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide Sato, Shinobu Ohzawa, Shinsuke Sota, Kojiro Sakamoto, Naoto Udo, Ayano Sueda, Shinji Matsuda, Tomoki Nagai, Takeharu Takenaka, Shigeori Front Chem Chemistry Potassium-sensing oligonucleotide, PSO, a conjugate of a quadruplex structure-forming oligonucleotide with a peptide incorporating a Förster Resonance Energy Transfer (FRET) chromophore pair, has been developed for fluorescent detection of potassium ion (K(+)) in aqueous medium. PSO 1 could be introduced into cells for real-time imaging of cytoplasmic K(+) concentrations. To perform fluorescent imaging of K(+) on the cell surface, we synthesized twelve PSO derivatives with different types of peptide types and lengths, and oligonucleotide sequences including thrombin-binding aptamer (TBA) sequences with FAM and TAMRA as a FRET chromophore pair, and evaluated their performance. 1 was shown to respond selectively to K(+), not to most ions present in vivo, and to show reciprocal fluorescence changes in response to K(+) concentration. For the peptide chains and oligonucleotide sequences examined in this study, the PSO derivatives had K (d) values for K(+) in the range of 5–30 mM. All PSO derivatives showed high K(+) selectivity even in the presence of excess Na(+). The PSO derivatives were successfully localized to the cell surface by biotinylated concanavalin A (ConA) or sulfo-NHS-biotin via streptavidin (StAv). Fluorescence imaging of extracellular K(+) upon addition of apoptosis inducers was successfully achieved by 1 localized to the cell surface. Frontiers Media S.A. 2022-07-08 /pmc/articles/PMC9306769/ /pubmed/35873036 http://dx.doi.org/10.3389/fchem.2022.922094 Text en Copyright © 2022 Sato, Ohzawa, Sota, Sakamoto, Udo, Sueda, Matsuda, Nagai and Takenaka. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Chemistry
Sato, Shinobu
Ohzawa, Shinsuke
Sota, Kojiro
Sakamoto, Naoto
Udo, Ayano
Sueda, Shinji
Matsuda, Tomoki
Nagai, Takeharu
Takenaka, Shigeori
Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide
title Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide
title_full Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide
title_fullStr Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide
title_full_unstemmed Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide
title_short Fluorescence Imaging of Extracellular Potassium Ion Using Potassium Sensing Oligonucleotide
title_sort fluorescence imaging of extracellular potassium ion using potassium sensing oligonucleotide
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9306769/
https://www.ncbi.nlm.nih.gov/pubmed/35873036
http://dx.doi.org/10.3389/fchem.2022.922094
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