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Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter

Chloride plays a major role in cellular homeostasis by regulating the lumenal pH of intracellular organelles. We have described a pH-independent, fluorescent chloride reporter called Clensor that has successfully measured resting chloride in organelles of living cells. Here, we describe the rational...

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Autores principales: Prakash, Ved, Saha, Sonali, Chakraborty, Kasturi, Krishnan, Yamuna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042475/
https://www.ncbi.nlm.nih.gov/pubmed/30050672
http://dx.doi.org/10.1039/c5sc04002g
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author Prakash, Ved
Saha, Sonali
Chakraborty, Kasturi
Krishnan, Yamuna
author_facet Prakash, Ved
Saha, Sonali
Chakraborty, Kasturi
Krishnan, Yamuna
author_sort Prakash, Ved
collection PubMed
description Chloride plays a major role in cellular homeostasis by regulating the lumenal pH of intracellular organelles. We have described a pH-independent, fluorescent chloride reporter called Clensor that has successfully measured resting chloride in organelles of living cells. Here, we describe the rational design of Clensor. Clensor integrates a chloride sensitive fluorophore called 10,10′-bis[3-carboxypropyl]-9,9′-biacridinium dinitrate (BAC) with the programmability, modularity and targetability available to nucleic acid scaffolds. We show that simple conjugation of BAC to a DNA backbone fails to yield a viable chloride-sensitive reporter. Fluorescence intensity and lifetime investigations on a series of BAC-functionalized structural variants yielded molecular insights that guided the rational design and successful realization of the chloride sensitive fluorescent reporter, Clensor. This study provides some general design principles that would aid the realization of diverse ion-sensitive nucleic acid reporters based on the sensing strategy of Clensor.
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spelling pubmed-60424752018-07-26 Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter Prakash, Ved Saha, Sonali Chakraborty, Kasturi Krishnan, Yamuna Chem Sci Chemistry Chloride plays a major role in cellular homeostasis by regulating the lumenal pH of intracellular organelles. We have described a pH-independent, fluorescent chloride reporter called Clensor that has successfully measured resting chloride in organelles of living cells. Here, we describe the rational design of Clensor. Clensor integrates a chloride sensitive fluorophore called 10,10′-bis[3-carboxypropyl]-9,9′-biacridinium dinitrate (BAC) with the programmability, modularity and targetability available to nucleic acid scaffolds. We show that simple conjugation of BAC to a DNA backbone fails to yield a viable chloride-sensitive reporter. Fluorescence intensity and lifetime investigations on a series of BAC-functionalized structural variants yielded molecular insights that guided the rational design and successful realization of the chloride sensitive fluorescent reporter, Clensor. This study provides some general design principles that would aid the realization of diverse ion-sensitive nucleic acid reporters based on the sensing strategy of Clensor. Royal Society of Chemistry 2016-03-01 2015-12-01 /pmc/articles/PMC6042475/ /pubmed/30050672 http://dx.doi.org/10.1039/c5sc04002g Text en This journal is © The Royal Society of Chemistry 2016 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0)
spellingShingle Chemistry
Prakash, Ved
Saha, Sonali
Chakraborty, Kasturi
Krishnan, Yamuna
Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter
title Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter
title_full Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter
title_fullStr Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter
title_full_unstemmed Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter
title_short Rational design of a quantitative, pH-insensitive, nucleic acid based fluorescent chloride reporter
title_sort rational design of a quantitative, ph-insensitive, nucleic acid based fluorescent chloride reporter
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042475/
https://www.ncbi.nlm.nih.gov/pubmed/30050672
http://dx.doi.org/10.1039/c5sc04002g
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