Enhanced recognition of cystic fibrosis W1282X DNA point mutation by chiral peptide nucleic acid probes by a surface plasmon resonance biosensor.

Abstract

Peptide nucleic acids (PNAs) containing an insert of three chiral monomers based on D-lysine ('chiral box') were synthesized and used as probes in Biospecific Interaction Analysis (BIA) for the recognition of DNA containing the W1282X point mutation of the cystic fibrosis gene. Hybridization experiments carried out in solution showed enhanced mismatch recognition when compared with the analogous achiral PNAs and oligonucleotides. The signal intensity was lower, but the selectivity of the Biacore response was found to be much higher than that observed with achiral PNAs. The newly designed chiral PNA probes were also found to hybridize with a 1:1 mixture of normal (N-W1282X) and mutated (M-W1282X) DNA oligomers immobilized on the biosensor, thus allowing discrimination not only between a normal and a mutated sequence (healthy/homozygous), but also between homo- and heterozygous individuals. These results suggest that 'chiral box' PNAs are potential powerful tools for the analysis of single point mutations of biological/biomedical relevance.

• Authors: Roberto Corradini, Giordana Feriotto, Stefano Sforza, Rosangela Marchelli, Roberto Gambari
• Journal: Journal of molecular recognition : JMR (J Mol Recognit) 2004 Jan-Feb Vol. 17 Issue 1 Pg. 76-84 ISSN: 0952-3499 [Print] England
• PMID: 14872540 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 2003 John Wiley & Sons, Ltd.

Chemical References

• Peptide Nucleic Acids
• Lysine

Topics

• Cystic Fibrosis (genetics)
• Humans
• Lysine (genetics)
• Mutation, Missense (genetics)
• Nucleic Acid Hybridization
• Peptide Nucleic Acids (analysis, chemical synthesis, chemistry, genetics)
• Point Mutation (genetics)
• Sensitivity and Specificity
• Surface Plasmon Resonance (instrumentation, methods)
• Transition Temperature