"It has also previously been shown that gold nanoparticles that are densely functionalized with DNA bind complementary DNA in a highly cooperative manner, resulting in a binding strength that is two orders-of-magnitude greater than that determined for analogous DNA strands that are not attached to a gold nanoparticle. This property has rendered nanoparticles particularly useful for DNA and protein diagnostic assays in addition to those uses described above.
"One class of oligonucleotides of interest are those that can detect a specific target with a recognition sequence. These types of structures, if introduced into living cells, are especially of interest for medical diagnosis, drug discovery, and developmental and molecular biology application. However, current delivery/transfection strategies lack the attributes required for their use such as 1) low toxicity, 2) high cellular uptake, and 3) provide resistance to enzymes that lead to false positive signals.
"Probes to visualize and detect intracellular RNA including those used for in situ staining (Femino et al., Science 280: 585-590, 1998; Kloosterman et al., Nat. Methods 3: 27-29, 2006), molecular beacons (Tyagi et al., 1996, Nat. Biotechnol. 14: 303-308; Sokol et al., 1998, Proc. Natl. Acad. Sci.
"Accordingly, while nanoparticle have been designed that can recognize targets with a high degree of specificity, it is difficult to detect a positive effect arising from the specific interaction, particularly with the sensitivity to detect such an interaction at the single cell level.
"Thus there exists a need in the art to develop materials which are capable of entering a cell to associate with a specific target and methods to detect and quantitate the resulting intracellular interaction."
Supplementing the background information on this patent, NewsRx reporters also obtained the inventors' summary information for this patent: "Provided here are methods of determining the intracellular concentration of a target molecule comprising the step of contacting the target molecule with a nanoparticle under conditions that allow association of the target molecule with the nanoparticle, the nanoparticle comprising a binding moiety specific for said target molecule, the binding moiety labeled with a marker, wherein the association of the target molecule and the nanoparticle results in detectable change in the marker, and wherein the change in the detectable marker is proportional to the intracellular concentration of said target molecule.
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