What Makes Thienoguanosine an Outstanding Fluorescent DNA Probe?

ABSTRACT: 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.

SUBMITTER: Kuchlyan J 

PROVIDER: S-EPMC7544670 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature