Detection of oligonucleotides by PNA-peptide conjugates recognizing the biarsenical fluorescein complex FlAsH-EDT2

Linda Piras; Concetta Avitabile; Luca Domenico D'Andrea; Michele Saviano; Alessandra Romanelli

doi:10.1016/j.bbrc.2017.09.064

Detection of oligonucleotides by PNA-peptide conjugates recognizing the biarsenical fluorescein complex FlAsH-EDT₂

Biochem Biophys Res Commun. 2017 Nov 4;493(1):126-131. doi: 10.1016/j.bbrc.2017.09.064. Epub 2017 Sep 15.

Authors

Linda Piras¹, Concetta Avitabile², Luca Domenico D'Andrea², Michele Saviano¹, Alessandra Romanelli³

Affiliations

¹ Institute of Crystallography (IC), CNR, Via Amendola 122/O, 70126, Bari, Italy.
² Institute of Biostructures and Bioimaging (IBB), CNR, via Mezzocannone 16, 80134, Napoli, Italy.
³ Department of Pharmacy, University of Naples "Federico II", via Mezzocannone 16, 80134, Napoli, Italy. Electronic address: alessandra.romanelli@unina.it.

PMID: 28919425
DOI: 10.1016/j.bbrc.2017.09.064

Abstract

We report the application of the arsenical complex FlAsH-EDT₂ for the identification of oligonucleotide sequences. We designed PNA sequences conjugated to either a tetracysteine motif and to split tetracysteine sequences, that are recognized by FlAsH. The effect of conjugation of the PNA to the tetracysteine peptide and RNA hybridization on the fluorescence of the arsenical complex has been investigated. The reconstitution of the tetracysteine motif, starting from 15-mer PNAs conjugated to split tetracysteine sequences and hybridized to a complementary oligonucleotide was also explored.

Keywords: Duplex; FlAsH-EDT(2); Hybridization; MYCN; Peptide nucleic acids; miR 17-5p.

MeSH terms

Fluoresceins / chemistry*
High-Throughput Nucleotide Sequencing / methods*
Oligonucleotide Array Sequence Analysis / methods*
Organometallic Compounds / chemistry*
Peptide Nucleic Acids / genetics*
Spectrometry, Fluorescence / methods*

Substances

4',5'-bis(1,3,2-dithioarsolan-2-yl)fluorescein
Fluoresceins
Organometallic Compounds
Peptide Nucleic Acids