A modular aldol approach for internal fluorescent molecular rotor chalcone surrogates for DNA biosensing applications

• Published in: Chemical science (Cambridge) 2023-05, Vol.14 (18), p.4832-4844
• Main Authors: Johnson, Ryan E, Murray, Makay T, Bycraft, Lucas J, Wetmore, Stacey D, Manderville, Richard A
• Format: Article
• Language: English
• Subjects: Aldehydes; Chemistry; Coupling (molecular); Fluorescence; Insertion; Molecular dynamics; Nucleic acids; Oligonucleotides
• ISSN: 2041-6520; 2041-6539
• DOI: 10.1039/d3sc00772c

Fluorescent molecular rotors (FMRs) are critical tools for probing nucleic acid structure and function. Many valuable FMRs have been incorporated into oligonucleotides, although the methods of doing so can be cumbersome. Development of synthetically simple, high yielding modular methods to fine-tune dye performance is crucial to expand the biotechnological applications of oligonucleotides. Herein, we report the utility of 6-hydroxy-indanone (6HI) with a glycol backbone to serve as a handle for on-strand aldehyde capture as a modular aldol approach for site-specific insertion of internal FMR chalcones. Aldol reactions with aromatic aldehydes containing N-donors proceed in high yield to create modified DNA oligonucleotides, which in the duplex match the stability of the fully paired canonical B-form with strong stacking interactions between the planar probe and the flanking base pairs, as evidenced by molecular dynamics (MD) simulations. The FMR chalcones possess remarkable quantum yields ( Φ fl up to 76%) in duplex DNA, coupled with large Stokes shifts (Δ ν up to 155 nm), light-up emissions ( I rel up to 60-fold) that span the visible region ( λ em 518-680 nm) with brightness up to 17 480 cm −1 M −1 . The library also contains a FRET pair and dual emission probes, suitable for ratiometric sensing. The ease of aldol insertion coupled with the excellent performance of the FMR chalcones permits their future wide-spread use. Insertion of an indanone handle into DNA oligonucleotides permits base-catalyzed aldol to create fluorescent molecular rotor (FMR) chalcones with turn-on fluorescence and brightness suitable for biosensing applications.