Quantification and spatio-temporal tracking of a target using a spherical nucleic acid (SNA)

Patent metadata

Field	Value
Publication number	US-10301622-B2
Application number	US-201415034005-A
Country	US
Kind code	B2
Filing date	Nov 4, 2014
Priority date	Nov 4, 2013
Publication date	May 28, 2019
Grant date	May 28, 2019

Abstract

The present invention relates to methods of detecting and tracking a target molecule using a nanoparticle wherein the nanoparticle comprises a polynucleotide that can specifically associate with the target molecule, and wherein the association results in a change in a detectable marker that can be measured after association with the target molecule.

First claim

What is claimed is:

1. A method comprising:
contacting a target polynucleotide with a composition comprising a nanoparticle under conditions that allow association of the target polynucleotide with the nanoparticle;
the nanoparticle comprising a first polynucleotide attached thereto, wherein a portion of the first polynucleotide comprises a sequence that is identical to a portion of the target polynucleotide;
the nanoparticle further comprising a second polynucleotide, wherein the second polynucleotide:
(i) comprises a marker;
(ii) is hybridized to the first polynucleotide; and
(iii) wherein hybridization of the second polynucleotide to the first polynucleotide results in an overhang of the second polynucleotide, wherein the overhang is from about 2 to about 30 nucleotides in length;

wherein association of the target polynucleotide and the nanoparticle results in:
(i) release of the second polynucleotide from the nanoparticle; and
(ii) association of the second polynucleotide and the target polynucleotide;

the association causing a detectable signal.

2. The method of claim 1 , wherein the position of the signal is determined.

3. The method of claim 1 , wherein the detectable signal is measured at time X and at time Y, wherein time Y is subsequent to time X.

4. The method of claim 3 , wherein the position of the signal is determined at time X and at time Y.

5. The method of claim 4 , wherein the change in position between time X and time Y is determined.

6. The method of claim 1 , wherein the detectable signal is measured in vitro.

7. The method of claim 1 , wherein the detectable signal is measured in a cell.

8. The method of claim 7 , wherein the cell is fixed and permeabilized.

9. The method of claim 1 , wherein the first polynucleotide and/or the second polynucleotide is DNA.

10. The method of claim 1 , wherein the first polynucleotide and/or the second polynucleotide is RNA.

11. The method of claim 1 , wherein the marker is quenched when the second polynucleotide comprising the marker is hybridized to the first polynucleotide.

12. The method of claim 1 , wherein the second polynucleotide comprises a marker which is a detectable label, wherein the marker is detectable only when the second polynucleotide is associated with the target polynucleotide.

13. The method of claim 1 , wherein the nanoparticle comprises a multiplicity of first polynucleotides and a multiplicity of second polynucleotides.

14. The method of claim 13 wherein at least one polynucleotide in the multiplicity of second polynucleotides associates with a different target polynucleotide than at least one other polynucleotide in the multiplicity of second polynucleotides.

15. The method of claim 1 , wherein the target polynucleotide is a non-coding RNA.

16. The method of claim 15 , wherein the non-coding RNA is a piwi-interacting RNA (piRNA).

17. The method of claim 1 , wherein the composition further comprises a therapeutic agent.

18. The method of claim 1 wherein the second polynucleotide hybridizes over the entire length of the first polynucleotide.

19. The method of claim 1 wherein the nanoparticle comprises about 10 second polynucleotides.

20. The method of claim 1 wherein the difference in melting temperature (T m ) between the first polynucleotide and the second polynucleotide is about 20-25° C.

21. The method of claim 1 wherein the nanoparticle comprises gold, silver copper, or platinum.

22. The method of claim 21 wherein the nanoparticle comprises gold.

Assignees

• Univ Northwestern

Inventors

• Mirkin Chad A
• Briley William E
• Randeria Pratik S
• Kim Nathaniel J

Classifications

• C12Q1/6823 Primary

  Release of bound markers · CPC title

• C12N2310/3517

  Marker; Tag · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• A61P35/00

  Antineoplastic agents · CPC title