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Snap-to-it probes: chelate-constrained nucleobase oligomers with enhanced binding specificity

Morgan, Joel R ; Lyon, Robert P ; Maeda, Dean Y ; Zebala, John A

Nucleic acids research, 2008-06, Vol.36 (11), p.3522-3530 [同儕審閱期刊]

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題名:
Snap-to-it probes: chelate-constrained nucleobase oligomers with enhanced binding specificity
著者: Morgan, Joel R ; Lyon, Robert P ; Maeda, Dean Y ; Zebala, John A
主題: Chemistry ; DNA - chemistry ; Fluorescent Dyes ; Metals - chemistry ; Nucleic Acid Conformation ; Nucleic Acid Denaturation ; Nucleic Acid Probes - chemistry ; Peptide Nucleic Acids - chemistry ; Temperature
所屬期刊: Nucleic acids research, 2008-06, Vol.36 (11), p.3522-3530
描述: We describe snap-to-it probes, a novel probe technology to enhance the hybridization specificity of natural and unnatural nucleic acid oligomers using a simple and readily introduced structural motif. Snap-to-it probes were prepared from peptide nucleic acid (PNA) oligomers by modifying each terminus with a coordinating ligand. The two coordinating ligands constrain the probe into a macrocyclic configuration through formation of an intramolecular chelate with a divalent transition metal ion. On hybridization with a DNA target, the intramolecular chelate in the snap-to-it probe dissociates, resulting in the probe 'snapping-to' and binding the target nucleic acid. Thermal transition analysis of snap-to-it probes with complementary and single-mismatch DNA targets revealed that the transition between free and target-bound probe conformations was a reversible equilibrium, and the intramolecular chelate provided a thermodynamic barrier to target binding that resulted in a significant increase in mismatch discrimination. A 4-6°C increase in specificity (ΔTm) was observed from snap-to-it probes bearing either terminal iminodiacetic acid ligands coordinated with Ni²⁺, or terminal dihistidine and nitrilotriacetic acid ligands coordinated with Cu²⁺. The difference in specificity of the PNA oligomer relative to DNA was more than doubled in snap-to-it probes. Snap-to-it probes labeled with a fluorophore-quencher pair exhibited target-dependent fluorescence enhancement upon binding with target DNA.
出版者: England: Oxford University Press
語言: 英文
識別號: ISSN: 0305-1048
EISSN: 1362-4962
DOI: 10.1093/nar/gkn219
PMID: 18448470
CODEN: NARHAD
資源來源: Directory of Open Access Journals

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Snap-to-it probes: chelate-constrained nucleobase oligomers with enhanced binding specificity

Morgan, Joel R ; Lyon, Robert P ; Maeda, Dean Y ; Zebala, John A

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