Steady-State Fluorescence and Lifetime Emission Study of pH-Sensitive Probes Based on i-motif Forming Oligonucleotides Single and Double Labeled with Pyrene

Cytosine-rich nucleic acids undergo pH-stimulated structural transitions leading to formation of an i-motif architecture at an acidic pH. Thus, i-motifs are good foundation for designing simple pH-sensitive fluorescent probes. We report here steady-state and time-resolved fluorescence studies of pyrene-labeled probes based on RET sequence: C(4)GC(4)GC(4)GC(4)TA (RET21), AC(4)GC(4)GC(4)GC(4)TA (RET21A) and C(4)GC(4)GC(4)GC(4)T (RET20). Comparative studies with single- and double-labeled i-motif probes were carried out. For each probe, we have measured fluorescence spectra and decays for emission wavelength of 390 nm over a wide range of pH (from 4.0 to 8.0). Effect of the oligonucleotide sequence and the number of pyrene labels on the spectral characteristics of probes were discussed.