Electronic Modifications of Fluorescent Cytidine Analogues Control Photophysics and Fluorescent Responses to Base Stacking and Pairing

• Published in: Chemistry : a European journal Vol. 25; no. 5; pp. 1249 - 1259
• Main Authors: Teppang, Kristine L., Lee, Raymond W., Burns, Dillon D., Turner, M. Benjamin, Lokensgard, Melissa E., Cooksy, Andrew L., Purse, Byron W.
• Format: Journal Article
• Language: English
• Published: WEINHEIM Wiley 24.01.2019; Wiley Subscription Services, Inc
• Subjects: Base stacking; biophysics; Chemistry; Chemistry, Multidisciplinary; Deoxyribonucleic acid; DNA; Dynamic structural analysis; Electrons; Fine structure; Fluorescence; Nucleoside analogs; nucleoside analogues; Nucleosides; Oligonucleotides; Oxidation; photochemistry; Physical Sciences; Science & Technology; Stacking; Ultrastructure; APPROXIMATION; NUCLEIC-ACID; ENVIRONMENT; NUCLEOSIDE; NUCLEOTIDES; SENSITIVE FLUORESCENT; ATOMS LI; fluorescence; photochemistry; DNA; nucleoside analogues; EXCHANGE; biophysics; GAUSSIAN-BASIS SETS
• ISSN: 0947-6539; 1521-3765
• DOI: 10.1002/chem.201803653

The rational design of fluorescent nucleoside analogues is greatly hampered by the lack of a general method to predict their photophysics, a problem that is especially acute when base pairing and stacking change fluorescence. To better understand these effects, a series of tricyclic cytidine (tC and tCO) analogues ranging from electron‐rich to electron‐deficient was designed and synthesized. They were then incorporated into oligonucleotides, and photophysical responses to base pairing and stacking were studied. When inserted into double‐stranded DNA oligonucleotides, electron‐rich analogues exhibit a fluorescence turn‐on effect, in contrast with the electron‐deficient compounds, which show diminished fluorescence. The magnitude of these fluorescence changes is correlated with the oxidation potential of nearest neighbor nucleobases. Moreover, matched base pairing enhances fluorescence turn‐on for the electron‐rich compounds, and it causes a fluorescence decrease for the electron‐deficient compounds. For the tCO compounds, the emergence of vibrational fine structure in the fluorescence spectra in response to base pairing and stacking was observed, offering a potential new tool for studying nucleic acid structure and dynamics. These results, supported by DFT calculations, help to rationalize fluorescence changes in the base stack and will be useful for selecting the best fluorescent nucleoside analogues for a desired application. Base stacking and pairing of fluorescent nucleosides such as DEA‐tC change their photophysical properties in ways that are strongly correlated with the electronic nature of neighboring bases.