POLYMER-BASED SIGNAL AMPLIFICATION FOR PROTEIN AND CELL DETECTION
The present invention relates generally to compositions, methods and kits for detection of a molecule of interest. The present invention is based, at least in part, on the design of nucleic acid oligonucleotides such that they can be used as molecular building blocks which can be assembled (polymerized) to form a dsDNA polymerization product which can be detected and further, can be disassembled (depolymerized) such that the polymerization product is no longer detectable. The invention can be performed multiple times on the same cell population, and is therefore useful for highly sensitive in situ detection of multiple biomarkers.
1 - 41 . (canceled)
42 . A kit comprising an initiator ssDNA oligonucleotide, at least one hairpin DNA oligonucleotide, and a depolymerization ssDNA oligonucleotide.
43 . The kit of claim 42 , wherein at least one hairpin DNA oligonucleotide is conjugated to a detectable molecule.
44 . The kit of claim 42 , wherein the initiator ssDNA oligonucleotide is linked to a molecule as a targeting complex.
45 . The kit of claim 42 , further comprising a ssDNA oligonucleotide conjugated to a detectable molecule.
46 . The kit of claim 45 , wherein the detectable molecule is selected from the group consisting of a quantum dot, a DNA-conjugated monomeric fluorophore and a DNA-conjugated polymeric fluorophore.
47 . The kit of claim 46 , wherein a DNA-conjugated fluorophore is selected from the group consisting of a succinimidyl ester activated fluorophore, Coumarin, FITC and TRITC.
48 . The kit of claim 42 , further comprising
a) a ssDNA oligonucleotide conjugated to biotin and
b) a streptavidin conjugated detectable molecule.
49 . The kit of claim 48 , wherein the streptavidin conjugated detectable molecule is a quantum dot.