Sequence-Selective Recognition of DNA by Strand Displacement with a Thymine-Substituted Polyamide

Nielsen, Peter E.; Egholm, Michael; Berg, Rolf H.; Buchardt, Ole

doi:10.1126/science.1962210

Cited by 3,109 publications

(2,290 citation statements)

References 26 publications

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“…6 where a 10-nucleobase target is readily probed in competition with reannealing of a 355-bp PCR amplicon. In fact, heating may not even be necessary since PNA, under certain conditions, hybridizes to DNA by strand invasion (21)(22)(23)(24)(25).…”

Section: Light-up Probes Are Suitable For Hybridization Assays In Hommentioning

confidence: 99%

Light-Up Probes: Thiazole Orange-Conjugated Peptide Nucleic Acid for Detection of Target Nucleic Acid in Homogeneous Solution

Svanvik

Westman

Wang³

et al. 2000

Analytical Biochemistry

237

147

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We have constructed light-up probes for nucleic acid detection. The light-up probe is a peptide nucleic acid (PNA) oligonucleotide to which the asymmetric cyanine dye thiazole orange (TO) is tethered. It combines the excellent hybridization properties of PNA and the large fluorescence enhancement of TO upon binding to DNA. When the PNA hybridizes to target DNA, the dye binds and becomes fluorescent. Free probes have low fluorescence, which may increase almost 50-fold upon hybridization to complementary nucleic acid. This makes the light-up probes particularly suitable for homogeneous hybridization assays, where separation of the bound and free probe is not necessary. We find that the fluorescence enhancement upon hybridization varies among different probes, which is mainly due to variations in free probe fluorescence. For eight probes studied the fluorescence quantum yield at 25°C in the unbound state ranged from 0.0015 to 0.08 and seemed to depend mainly on the PNA sequence. The binding of the light-up probes to target DNA is highly sequence specific and a single mismatch in a 10-mer target sequence was readily identified.

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Section: Light-up Probes Are Suitable For Hybridization Assays In Hommentioning

confidence: 99%

Light-Up Probes: Thiazole Orange-Conjugated Peptide Nucleic Acid for Detection of Target Nucleic Acid in Homogeneous Solution

Svanvik

Westman

Wang³

et al. 2000

Analytical Biochemistry

237

147

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“…1,2 Their neutral pseudopeptide backbone is made of N- (2-aminoethyl)glycine units which are ligated via a methylene carbonyl to the four nucleobases ( Figure 1). 1 Single-stranded (ss) PNAs bind to complementary ssDNA, 1, 3 RNA 3 or PNA 4 sequences obeying the Watson-Crick base-pairing rules. Hybridisation may occur in a parallel, or more favourably, in an antiparallel fashion.…”

Section: Introductionmentioning

confidence: 99%

Metal-containing peptide nucleic acid conjugates

2011

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Peptide Nucleic Acids (PNAs) are non-natural DNA/RNA analogues with favourable physicochemical properties and promising applications. Discovered nearly 20 years ago, PNAs have recently re-gained quite a lot of attention. In this Perspective article, we discuss the latest advances on the preparation and utilisation of PNA monomers and oligomers containing metal complexes. These metalconjugates have found applications in various research fields such as in the sequence-specific detection of nucleic acids, in the hydrolysis of nucleic acids and peptides, as radioactive probes or as modulators of PNA˙DNA hybrid stability, and last but not least as probes for molecular and cell biology.

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“…The stability and specificity of the binding between DNA or RNA targets and supportbound probe molecules are crucial factors for the quality and robustness of the analyses. PNA is an entirely synthetic DNA mimic that was invented by Peter Nielsen et al [1]. Its particular chemical properties make it suitable for use in bioassays.…”

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confidence: 99%

“…Its particular chemical properties make it suitable for use in bioassays. In PNA, the negatively charged (deoxy-) ribose-phosphate backbone of nucleic acids is replaced by an uncharged N-(2-aminoethyl)-glycine scaffold, to which the nucleobases are attached via a methylene carbonyl linker [1,2] (Figure 1). Because all intramolecular distances and the configuration of the nucleobases are similar to those in natural DNA molecules, specific hybridization occurs between PNA and cDNA or RNA sequences.…”

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confidence: 99%