Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores
AssigneeThe Regents of the University of California
InventorsGuillermo C. BAZAN, Brent S. GAYLORD, Shu WANG
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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 polynucleotide complementary to the target polynucleotide. The sensor polynucleotide comprises a signaling chromophore to receive energy from the excited multichromophore and increase emission 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.
Claims20
An assay method comprising:
- providing a sample that is suspected of containing a target polynucleotide;
- providing a polycationic multichromophore that upon excitation is capable of transferring energy to a signaling chromophore;
- providing an anionic sensor polynucleotide that is single-stranded and is complementary to the target polynucleotide, said sensor polynucleotide conjugated to the signaling chromophore, wherein said sensor polynucleotide interacts with the multichromophore and emitted light can be produced from the signaling chromophore upon excitation of the multichromophore in the absence of target polynucleotide, and wherein a greater amount of emitted light is produced from the signaling chromophore upon excitation of the multichromophore in the presence of target polynucleotide;
- contacting the sample with the sensor polynucleotide and the multichromophore in a solution under conditions in which the sensor polynucleotide can hybridize to the target polynucleotide, if present;
- applying a light source to the solution that can excite the multichromophore;
- and
- detecting whether the light emitted from the signaling chromophore is increased in the presence of sample.
The method of claim 30, wherein the substrate is conjugated to a plurality of different sensor polynucleotides having corresponding different sequences, wherein each of said different sensor polynucleotides can selectively hybridize to a corresponding different target polynucleotide.
A polynucleotide sensing solution comprising:
- a signaling chromophore;
- and
- a polycationic multichromophore that is capable of transferring energy to the signaling chromophore upon excitation when brought into proximity thereto, wherein a greater amount of energy can be produced from the signaling chromophore in the presence of the polynucleotide being sensed when the multichromophore is excited.
A sensing complex comprising:
- a sensor polynucleotide that is single-stranded and is complementary to a target polynucleotide, said sensor polynucleotide attached to a signaling chromophore;
- a polycationic multichromophore that is capable of transferring energy to the signaling chromophore upon excitation when brought into proximity thereto, wherein said sensor polynucleotide interacts with the multichromophore and emitted light can be produced from the signaling chromophore upon excitation of the multichromophore in the absence of target polynucleotide, and wherein a greater amount of emitted light is produced from the signaling chromophore upon excitation of the multichromophore in the presence of target polynucleotide.
Dependencies are inferred from the claim text.