Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores
First Claim
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1. An assay method comprising:
- providing a sample that is suspected of containing a target polynucleotide;
providing a polycationic multichromophore that electrostatically interacts with the target polynucleotide and upon excitation is capable of transferring energy to a signaling chromophore;
providing a sensor peptide nucleic acid (PNA) that is single-stranded and is complementary to the target polynucleotide, said sensor PNA conjugated to the signaling chromophore;
contacting the sample with the sensor PNA and the multichromophore in a solution under conditions in which the sensor PNA can hybridize to the target polynucleotide, if present;
applying a light source to the solution that can excite the multichromophore; and
detecting whether light is emitted from the signaling chromophore.
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Abstract
Methods, compositions and articles of manufacture for assaying a sample for a target polynucleotide are provided. A sample suspected of containing the target polynucleotide is contacted with a polycationic multichromophore and a sensor PNA complementary to the target polynucleotide. The sensor PNA comprises a signaling chromophore to absorb energy from the excited multichromophore and emit light in the presence of the target polynucleotide. The methods can be used in multiplex form. Kits comprising reagents for performing such methods are also provided.
49 Citations
30 Claims
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1. An assay method comprising:
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providing a sample that is suspected of containing a target polynucleotide;
providing a polycationic multichromophore that electrostatically interacts with the target polynucleotide and upon excitation is capable of transferring energy to a signaling chromophore;
providing a sensor peptide nucleic acid (PNA) that is single-stranded and is complementary to the target polynucleotide, said sensor PNA conjugated to the signaling chromophore;
contacting the sample with the sensor PNA and the multichromophore in a solution under conditions in which the sensor PNA can hybridize to the target polynucleotide, if present;
applying a light source to the solution that can excite the multichromophore; and
detecting whether light is emitted from the signaling chromophore. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 25, 26, 27, 28, 29, 30)
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23. A polynucleotide sensing solution comprising:
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a sensor peptide nucleic acid (PNA) that is single-stranded and is complementary to a target polynucleotide, said sensor PNA attached to a signaling chromophore;
a polycationic multichromophore that can electrostatically interact with the phosphate backbone of the target polynucleotide and is capable of transferring energy to the signaling chromophore upon excitation when brought into proximity thereto upon hybridization of the sensor PNA to the target polynucleotide.
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24. A kit for assaying a sample for a target polynucleotide comprising:
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a sensor peptide nucleic acid (PNA) that is single-stranded and is complementary to the target polynucleotide, said sensor PNA conjugated to a signaling chromophore; and
a polycationic multichromophore that can electrostatically interact with the phosphate backbone of the target polynucleotide and is capable of transferring energy to the signaling chromophore upon excitation when brought into proximity thereto upon hybridization of the sensor PNA to the target polynucleotide.
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Specification