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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
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High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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Optokinetically Encoded Nanoprobe-Based Multiplexing Strategy for MicroRNA Profiling.

Sungi Kim1, Jeong-Eun Park1, Woosung Hwang1

  • 1Department of Chemistry, Seoul National University , Seoul 08826, South Korea.

Journal of the American Chemical Society
|February 10, 2017
PubMed
Summary
This summary is machine-generated.

This study introduces a novel multiplexed molecular detection strategy using optokinetically coded nanoprobes for simultaneous, quantitative microRNA profiling. The assay enables rapid and specific detection of multiple microRNA targets, aiding in cancer diagnostics.

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Molecular Biology

Background:

  • Multiplexed analysis of interacting molecules is crucial for understanding biological systems, especially for microRNA (miRNA) profiling in cell biology, biosensing, and cancer diagnostics.
  • Accurate cancer diagnosis requires highly specific detection of multiple miRNA sequences simultaneously.

Purpose of the Study:

  • To develop a multiplexed molecular detection strategy for reliable and quantitative profiling of multiple microRNA targets.
  • To demonstrate the capability of the developed assay for potential application in cancer diagnostics.

Main Methods:

  • Utilized optokinetically (OK) coded nanoprobes (NPs) with distinct optical signals (red, green, blue) tethered to a supported lipid bilayer (SLB).
  • Employed dark-field microscopy (DFM) for in situ single-particle monitoring and normalized RGB analysis of combinatorial NP assemblies.
  • Developed the OK-nanoprobe-lipid bilayer (OK-NLB) assay for differentiating and quantifying multiple miRNA targets.

Main Results:

  • The OK-NLB assay successfully differentiated and quantified 9 different miRNA targets in a single sample.
  • Achieved simultaneous detection of multiple miRNA targets with high quantitation and specificity within 1 hour.
  • Validated the assay using single-base mismatched experiments and HeLa cell-extracted total RNA, showing comparable results to quantitative reverse transcription polymerase chain reaction (qRT-PCR).

Conclusions:

  • The OK-NP-based assay provides a powerful tool for multiplexed molecular detection with high sensitivity and specificity.
  • This assay has significant potential for advancing miRNA-based diagnostics, particularly for early cancer detection.