Anu Kiviniemi

Synthesis of fluorine-labeled peptide nucleic acid building blocks as sensors for the 19F NMR spectroscopic detection of different hybridization modes.

Peptide nucleic acid (PNA) building blocks, bearing a fluorine sensor at C-5 of the uracil base [viz. trifluoromethyl and 3,3-bis(trifluoromethyl)-4,4,4-trifluorobut-1-ynyl], were synthesized and incorporated to a PNA strand, and their applicability for the monitoring of different hybridization modes by (19)F NMR spectroscopy was studied. Both sensors gave unique (19)F resonance shifts in NMR when the PNA was targeted to a complementary antiparallel DNA, antiparallel RNA, parallel DNA, and parallel RNA. The 5-trifluoromethyl-derived sensor was additionally applied for the monitoring of interconversions from a parallel DNA/PNA complex to an antiparallel RNA/PNA complex and from a PNA/PNA complex to two DNA/PNA complexes (i.e., double-duplex invasion).

Oligonucleotides bearing pentaerythritol-derived mass tags

Five different di-O-alkylated pentaerythritol phosphoramidite building blocks (2a–e) were synthesized and introduced into oligonucleotides to obtain mass-tagged probes (6a–f) useful in RNA transcription profiling. These non-nucleosidic mass tags allow categorization of oligonucleotide probes having identical nucleoside content and, hence, identification of the probe hybridized to RNA by mass spectrometry analysis. Hybridization properties of the oligonucleotide conjugates were elucidated by melting temperature measurements.