DNA detection using water-soluble conjugated polymers and peptide nucleic acid probes

The light-harvesting properties of cationic conjugated polymers are used to sensitize the emission of a dye on a specific peptide nucleic acid (PNA) sequence for the purpose of homogeneous, “real-time” DNA detection. Signal transduction is controlled by hybridization of the neutral PNA probe and the negative DNA target. Electrostatic interactions bring the hybrid complex and cationic polymer within distances required for Förster energy transfer. Conjugated polymer excitation provides fluorescein emission >25 times higher than that obtained by exciting the dye, allowing detection of target DNA at concentrations of 10 pM with a standard fluorometer. A simple and highly sensitive assay with optical amplification that uses the improved hybridization behavior of PNA/DNA complexes is thus demonstrated.

[1]  A. Heeger,et al.  Energy Transfer in Mixtures of Water- Soluble Oligomers: Effect of Charge, Aggregation, and Surfactant Complexation , 2002 .

[2]  H. Stender,et al.  PNA for rapid microbiology. , 2002, Journal of microbiological methods.

[3]  A. Heeger,et al.  Water-soluble conjugated oligomers: effect of chain length and aggregation on photoluminescence-quenching efficiencies. , 2001, Journal of the American Chemical Society.

[4]  H. Yowanto,et al.  Electronic detection of nucleic acids: a versatile platform for molecular diagnostics. , 2001, The Journal of molecular diagnostics : JMD.

[5]  N. Schork,et al.  Single nucleotide polymorphisms and the future of genetic epidemiology , 2000, Clinical genetics.

[6]  Joseph Wang SURVEY AND SUMMARY From DNA biosensors to gene chips , 2000 .

[7]  D. Moses,et al.  Photoluminescence of water-soluble conjugated polymers : Origin of enhanced Quenching by charge transfer , 2000 .

[8]  T. Swager,et al.  Conjugated polymer-based chemical sensors. , 2000, Chemical reviews.

[9]  K. Ulbrich,et al.  Polyelectrolyte vectors for gene delivery: influence of cationic polymer on biophysical properties of complexes formed with DNA. , 1999, Bioconjugate chemistry.

[10]  F. Wudl,et al.  Highly sensitive biological and chemical sensors based on reversible fluorescence quenching in a conjugated polymer. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[11]  A. Heeger,et al.  Time-resolved Förster energy transfer in polymer blends , 1999 .

[12]  P. Nielsen,et al.  Applications of peptide nucleic acids. , 1999, Current opinion in biotechnology.

[13]  Harada,et al.  Chain length recognition: core-shell supramolecular assembly from oppositely charged block copolymers , 1999, Science.

[14]  P. Nielsen,et al.  A formula for thermal stability (Tm) prediction of PNA/DNA duplexes. , 1998, Nucleic acids research.

[15]  K. Balakin,et al.  Conjugates of oligonucleotides with polyaromatic fluorophores as promising DNA probes. , 1998, Biosensors & bioelectronics.

[16]  A. Kabanov,et al.  Self-assembling complexes for gene delivery : from laboratory to clinical trial , 1998 .

[17]  Alonso Castro,et al.  Single-Molecule Detection of Specific Nucleic Acid Sequences in Unamplified Genomic DNA , 1997 .

[18]  C. Pichot,et al.  Adsorption of Single-Stranded DNA Fragments onto Cationic Aminated Latex Particles , 1997 .

[19]  L. Betts,et al.  A Nucleic Acid Triple Helix Formed by a Peptide Nucleic Acid-DNA Complex , 1995, Science.

[20]  V V Demidov,et al.  Stability of peptide nucleic acids in human serum and cellular extracts. , 1994, Biochemical pharmacology.

[21]  Peter E. Nielsen,et al.  PNA hybridizes to complementary oligonucleotides obeying the Watson–Crick hydrogen-bonding rules , 1993, Nature.

[22]  J. Bisi,et al.  Antisense and antigene properties of peptide nucleic acids. , 1992, Science.

[23]  D. E. Wolf,et al.  Detection of nucleic acid hybridization by nonradiative fluorescence resonance energy transfer. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[24]  T. Julian,et al.  Permeability characteristics of calcium alginate films , 1988 .

[25]  J. Lepecq,et al.  A fluorescent complex between ethidium bromide and nucleic acids. Physical-chemical characterization. , 1967, Journal of molecular biology.