Methods and compositions for detection and analysis of polynucleotide-binding protein interactions 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 interacts with the target polynucleotide and upon excitation is capable of transferring energy to a signaling chromophore;
providing a sensor polynucleotide binding protein (PBP) that can bind to the target polynucleotide, said sensor PBP conjugated to the signaling chromophore;
contacting the sample with the sensor PBP and the multichromophore in a solution under conditions in which the sensor PBP can bind 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 PBP that can bind to the target polynucleotide. The sensor PBP 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.
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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 interacts with the target polynucleotide and upon excitation is capable of transferring energy to a signaling chromophore;
providing a sensor polynucleotide binding protein (PBP) that can bind to the target polynucleotide, said sensor PBP conjugated to the signaling chromophore;
contacting the sample with the sensor PBP and the multichromophore in a solution under conditions in which the sensor PBP can bind 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, 23, 24, 27, 28, 29, 30)
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25. A polynucleotide sensing solution comprising:
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a sensor polynucleotide binding protein (PBP) that binds to a target polynucleotide, said sensor PBP attached to a signaling chromophore;
a polycationic multichromophore that can electrostatically interact with the target polynucleotide and is capable of transferring energy to the signaling chromophore upon excitation when brought into proximity thereto upon binding of the sensor PBP to the target polynucleotide.
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26. A kit for assaying a sample for a target polynucleotide comprising:
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a sensor PBP that can bind to the target polynucleotide;
a polycationic multichromophore that can electrostatically interact with the target polynucleotide and is capable of transferring energy to the signaling chromophore upon excitation when brought into proximity thereto upon binding of the sensor PBP to the target polynucleotide;
a housing for retaining the sensor PBP and multichromophore; and
instructions provided with said housing that describe how to use the components of the kit to assay the sample for the target polynucleotide.
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Specification