Targeting a KRAS i-motif forming sequence by unmodified and gamma-modified peptide nucleic acid oligomers

January 31, 2023

Title

Targeting a KRAS i-motif forming sequence by unmodified and gamma-modified peptide nucleic acid oligomers

Author

Srijani Sarkar, Gabriela Colón-Roura, Alexander Pearse, Bruce A. Armitage

Year

2022

Journal

Biopolymers

Abstract

Growing interest in i-motif DNA as a transcriptional regulatory element motivates development of synthetic molecules capable of targeting these structures. In this study, we designed unmodified peptide nucleic acid (PNA) and gamma-modified PNA (γPNA) oligomers complementary to an i-motif forming sequence derived from the promoter of the KRAS oncogene. Biophysical techniques such as circular dichroism (CD) spectroscopy, CD melting, and fluorescence spectroscopy demonstrated the successful invasion of the i-motif by PNA and γPNA. Both PNA and γPNA showed very strong binding to the target sequence with high thermal stability of the resulting heteroduplexes. Interestingly fluorescence and CD experiments indicated formation of an intermolecular i-motif structure via the overhangs of target-probe heteroduplexes formed by PNA/γPNA invasion of the intramolecular i-motif. Targeting promoter i-motif forming sequences with high-affinity oligonucleotide mimics like γPNAs may represent a new approach for inhibiting KRAS transcription, thereby representing a potentially useful anti-cancer strategy.

Instrument

J-1500

Keywords

KRAS, i-motif, sequence, gamma-modified, peptide, nucleic acid, oligomers