Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores

• US 8,841,072 B2
• Filed: 11/21/2013
• Issued: 09/23/2014
• Est. Priority Date: 08/26/2002
• Status: Expired

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Bazan, Guillermo C., Gaylord, Brent S., Wang, Shu
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US14/086,532
• Publication Number:
  US 20140141428A1
• Time in Patent Office: 306 Days
• Field of Search: 435/6.1, 536/23.1, 536/26.6
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2523/313 Irradiation, e.g. UV irradiation
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2565/107 Alteration in the property of hybridised versus free label oligonucleotides
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

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.

Claims

1. A light harvesting multichromophore system, the system comprising

a) a signaling chromophore;

and

b) a water-soluble conjugated polymer comprising a delocalized electronic structure, wherein the polymer can transfer energy from its excited state to the signaling chromophore to provide a greater than 4 fold increase in fluorescence emission from the signaling chromophore than can be achieved by direct excitation.

2. The light harvesting multichromophore system of claim 1, wherein the polymer can transfer energy from its excited state to a signaling chromophore to provide an 8 fold increase in fluorescence emission from the signaling chromophore than can be achieved by direct excitation.

3. The light harvesting multichromophore system of claim 1, wherein the signaling chromophore comprises a fluorescent dye.

4. The light harvesting multichromophore system of claim 1, wherein the conjugated polymer has the structure

5. The light harvesting multichromophore system of claim 1, wherein the system further comprises a substrate.

6. The light harvesting multichromophore system of claim 1, wherein the system further comprises a second signaling chromophore.

7. The light harvesting multichromophore system according to claim 6, wherein the second signaling chromophore comprises a fluorescent dye.

8. The light harvesting multichromophore system according to claim 6, wherein the second signaling chromophore is attached to a substrate.

9. The light harvesting multichromophore system according to claim 1, further comprises an excitation source.

10. The light harvesting multichromophore system according to claim 9, wherein the excitation source comprises a laser.

11. The light harvesting multichromophore system according to claim 1, wherein the system further comprises a detector.

12. The light harvesting multichromophore system according to claim 11, wherein the detector comprises a fluorometer.