MicroRNAs (miRNAs) have recently emerged as promising biomarkers for cancer profiling with better performance than messenger RNA. Unfortunately, traditional miRNA analysis techniques lack quantitation, multiplexing, and/or throughput, and have clinically impractical sample preparation and assay workflows. This represents a major hurdle in transforming miRNA signatures from tumor cells to the clinical diagnosis and stratification of cancers. Consequently, there is a technological gap for miRNA probing technologies with simple, translatable workflows that can quantitatively resolve heterogeneity in cell populations. To address this gap, <u>we developed a miRNA analysis platform with the capacity to perform parallel, multiplex, and sensitive miRNA measurements in arrays of isolated nanoliter wells from raw cells without prior sample preparation (i.e. nucleic acid extraction)</u>.<br/><br/>Prior work showed the use of hydrogel particles functionalized with DNA probes for quantitative miRNA assays from raw cells. While robust, this particle-based assay requires samples of ~1000 cells in 100 μL reactors, thus limiting the sensitivity and scalability to assess heterogeneity in cell populations. Given these limitations, we developed the nanoliter well array approach capable of performing &gt;100 parallel assays with simple and robust assay workflows (and not needing sample preparation). The well reactors contain hydrogel posts each functionalized with different DNA probes for multiplex measurements. By reducing the reactor volumes and sensing surface, the well-based approach has ~50x higher sensitivity (0.025 attomoles miRNA, &lt;20 cells per reactor) without relying on signal amplification.

Multiplex microRNA assays from raw cells in isolated nanoliter well arrays

256th National Meeting (2018)

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