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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 the pre-miRNA...
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Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer
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Mutations in microRNA-128-2-3p identified with amplification-free hybridization assay.

Sofie Slott1, Cecilie Schiøth Krüger-Jensen1, Izabela Ferreira da Silva2,3,4

  • 1Department of Chemistry, Technical University of Denmark, Kgs Lyngby, Denmark.

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|August 22, 2023
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Summary

This study presents a new method to detect mutated microRNAs in plasma. The assay identifies specific mutations in microRNA-128-2-3p linked to inflammatory colitis and colorectal cancer.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Medical Diagnostics

Background:

  • MicroRNAs (miRNAs) are crucial regulators of gene expression.
  • Mutations in miRNAs, particularly single-nucleotide polymorphisms (SNPs), can alter their function.
  • Detecting miRNA mutations in accessible biofluids like plasma is vital for disease diagnosis.

Purpose of the Study:

  • To develop a quantitative, amplification-free method for detecting mutated microRNAs in human plasma.
  • To validate the assay's specificity and sensitivity using a Peyrard-Bishop model.
  • To investigate the correlation of specific miRNA mutations with inflammatory colitis and colorectal cancer.

Main Methods:

  • Design of specific oligonucleotides based on the Peyrard-Bishop model for accurate target:probe recognition.
  • Development of an amplification-free tandem bead-based hybridization assay.
  • Quantitative detection of single-nucleotide polymorphism (SNP) mismatch profiles in microRNA-128-2-3p.

Main Results:

  • The assay achieved a limit of detection of 2.2 pM.
  • Accurate prediction of target:probe recognition affinity and specificity was demonstrated.
  • Identification of terminal mutations in microRNA-128-2-3p.
  • Positive correlation found between identified mutations and inflammatory colitis/colorectal cancer.

Conclusions:

  • A novel, sensitive, and specific assay for quantitative detection of mutated miRNAs in plasma has been developed.
  • The assay can identify clinically relevant miRNA mutations.
  • The findings suggest potential utility of these miRNA mutations as biomarkers for inflammatory colitis and colorectal cancer.