biostudies-literatureKuchlyan JNvidiaAgence Nationale de la RechercheCentre National de la Recherche ScientifiqueInstitut Universitaire de FranceNational Institutes of HealthLabexNIGMS NIH HHSMinisterio de Econom?a y Competitividad16999-17014https://www.ebi.ac.uk/biostudies/studies/S-EPMC7544670biostudies-literatureUnknown142(40)Thienoguanosine (^thG) is an isomorphic guanosine (G) surrogate that almost perfectly mimics G in nucleic acids. To exploit its full potential and lay the foundation for future applications, 20 DNA duplexes, where the bases facing and neighboring ^thG were systematically varied, were thoroughly studied using fluorescence spectroscopy, molecular dynamics simulations, and mixed quantum mechanical/molecular mechanics calculations, yielding a comprehensive understanding of its photophysics in DNA. In matched duplexes, ^thG's hypochromism was larger for flanking G/C residues but its fluorescence quantum yield (QY) and lifetime values were almost independent of the flanking bases. This was attributed to high duplex stability, which maintains a steady orientation and distance between nucleobases, so that a similar charge transfer (CT) mechanism governs the photophysics of ^thG independently of its flanking nucleobases. ^thG can therefore replace any G residue in matched duplexes, while always maintaining similar photophysical features. In contrast, the local destabilization induced by a mismatch or an abasic site restores a strong dependence of ^thG's QY and lifetime values on its environmental context, depending on the CT route efficiency and solvent exposure of ^thG. Due to this exquisite sensitivity, ^thG appears ideal for monitoring local structural changes and single nucleotide polymorphism. Moreover, ^thG's dominant fluorescence lifetime in DNA is unusually long (9-29 ns), facilitating its selective measurement in complex media using a lifetime-based or a time-gated detection scheme. Taken together, our data highlight ^thG as an outstanding emissive substitute for G with good QY, long fluorescence lifetimes, and exquisite sensitivity to local structural changes.Journal of the American Chemical SocietyWhat Makes Thienoguanosine an Outstanding Fluorescent DNA Probe?PMC7544670GM 069773R01 GM069773CTQ2016-76061-PMori MImprota RBoudier CMartinez-Fernandez LTor YGavvala KCiaco SMely YKuchlyan JRichert LDidier PfalseWhat Makes Thienoguanosine an Outstanding Fluorescent DNA Probe?Thienoguanosine (^thG) is an isomorphic guanosine (G) surrogate that almost perfectly mimics G in nucleic acids. To exploit its full potential and lay the foundation for future applications, 20 DNA duplexes, where the bases facing and neighboring ^thG were systematically varied, were thoroughly studied using fluorescence spectroscopy, molecular dynamics simulations, and mixed quantum mechanical/molecular mechanics calculations, yielding a comprehensive understanding of its photophysics in DNA. In matched duplexes, ^thG's hypochromism was larger for flanking G/C residues but its fluorescence quantum yield (QY) and lifetime values were almost independent of the flanking bases. This was attributed to high duplex stability, which maintains a steady orientation and distance between nucleobases, so that a similar charge transfer (CT) mechanism governs the photophysics of ^thG independently of its flanking nucleobases. ^thG can therefore replace any G residue in matched duplexes, while always maintaining similar photophysical features. In contrast, the local destabilization induced by a mismatch or an abasic site restores a strong dependence of ^thG's QY and lifetime values on its environmental context, depending on the CT route efficiency and solvent exposure of ^thG. Due to this exquisite sensitivity, ^thG appears ideal for monitoring local structural changes and single nucleotide polymorphism. Moreover, ^thG's dominant fluorescence lifetime in DNA is unusually long (9-29 ns), facilitating its selective measurement in complex media using a lifetime-based or a time-gated detection scheme. Taken together, our data highlight ^thG as an outstanding emissive substitute for G with good QY, long fluorescence lifetimes, and exquisite sensitivity to local structural changes.2020-01-01T00:00:00Z2020 Oct2024-02-15T19:58:43.543Z2022-02-11T11:50:03.703ZS-EPMC75446703291555810.1021/jacs.0c06165