Fmoc Solid-Phase FIT-PNA Synthesis-Manual and Automated Methodologies

Peptide nucleic acids (PNA) [1] are synthetic DNA analogs that are used as DNA/RNA mimics for diagnostic [2, 3] and therapeutic purposes [2-5]. One sensing strategy is the use of fluorogenic probes. One such probe is forced intercalation (FIT) PNA, which has a surrogate base as a cyanine dye (e.g., BisQ, TO) that replaces one of the natural bases in the PNA sequence. FIT-PNAs fluoresce after DNA/RNA hybridization, and display sensitivity towards single base mismatches [6]. These, combined with other desirable features, render them highly attractive as RNA/DNA sensors.Herein, we describe the procedure for FIT-PNA synthesis, both by manual and automated methodologies.