2 research outputs found
Scanometric MicroRNA Array Profiling of Prostate Cancer Markers Using Spherical Nucleic Acid–Gold Nanoparticle Conjugates
We report the development of a novel Scanometric MicroRNA
(Scano-miR)
platform for the detection of relatively low abundance miRNAs with
high specificity and reproducibility. The Scano-miR system was able
to detect 1 fM concentrations of miRNA in serum with single nucleotide
mismatch specificity. Indeed, it provides increased sensitivity for
miRNA targets compared to molecular fluorophore-based detection systems,
where 88% of the low abundance miRNA targets could not be detected
under identical conditions. The application of the Scano-miR platform
to high density array formats demonstrates its utility for high throughput
and multiplexed miRNA profiling from various biological samples. To
assess the accuracy of the Scano-miR system, we analyzed the miRNA
profiles of samples from men with prostate cancer (CaP), the most
common noncutaneous malignancy and the second leading cause of cancer
death among American men. The platform exhibits 98.8% accuracy when
detecting deregulated miRNAs involved in CaP, which demonstrates its
potential utility in profiling and identifying clinical and research
biomarkers
Multiplexed Nanoflares: mRNA Detection in Live Cells
We report the development of the multiplexed nanoflare,
a nanoparticle agent that is capable of simultaneously detecting two
distinct mRNA targets inside a living cell. These probes are spherical nucleic acid (SNA) gold nanoparticle (Au NP) conjugates consisting of densely packed and highly oriented oligonucleotide sequences, many of which are hybridized to a reporter with a distinct fluorophore
label and each complementary to its corresponding mRNA target. When
multiplexed nanoflares are exposed to their targets, they provide
a sequence specific signal in both extra- and intracellular environments.
Importantly, one of the targets can be used as an internal control,
improving detection by accounting for cell-to-cell variations in nanoparticle
uptake and background. Compared to single-component nanoflares, these
structures allow one to determine more precisely relative mRNA levels
in individual cells, improving cell sorting and quantification