Lighting Up DNA with the Environment‐Sensitive Bright Adenine Analogue qAN4
| Autor: | Tom Brown, Marcus Wilhelmsson, Anders Foller Füchtbauer, Sangamesh Sarangamath, Mattias Bood, Moa Sandberg Wranne, Henrik Gradén, Morten Grøtli, Afaf H. El-Sagheer |
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| Rok vydání: | 2020 |
| Předmět: |
Circular dichroism
Phosphoramidite Fluorophore Molecular Structure 010405 organic chemistry Chemistry Oligonucleotide Base pair Adenine DNA General Chemistry 010402 general chemistry Photochemistry 01 natural sciences 0104 chemical sciences Thymine chemistry.chemical_compound Förster resonance energy transfer |
| Zdroj: | ChemPlusChem. 85:319-326 |
| ISSN: | 2192-6506 |
| Popis: | The fluorescent adenine analogue qAN4 was recently shown to possess promising photophysical properties, including a high brightness as a monomer. Here we report the synthesis of the phosphoramidite of qAN4 and its successful incorporation into DNA oligonucleotides using standard solid-phase synthesis. Circular dichroism and thermal melting studies indicate that the qAN4-modification has a stabilizing effect on the B-form of DNA. Moreover, qAN4 base-pairs selectively with thymine with mismatch penalties similar to those of mismatches of adenine. The low energy absorption band of qAN4 inside DNA has its peak around 358 nm and the emission in duplex DNA is partly quenched and blue-shifted (ca. 410 nm), compared to the monomeric form. The spectral properties of the fluorophore also show sensitivity to pH; a property that may find biological applications. Quantum yields in single-stranded DNA range from 1-29 % and in duplex DNA from 1-7 %. In combination with the absorptive properties, this gives an average brightness inside duplex DNA of 275 M-1 cm-1 , more than five times higher than the most used environment-sensitive fluorescent base analogue, 2-aminopurine. Finally, we show that qAN4 can be used to advantage as a donor for interbase FRET applications in combination with adenine analogue qAnitro as an acceptor. |
| Databáze: | OpenAIRE |
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