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Related Concept Videos

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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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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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Updated: Nov 18, 2025

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method
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MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method

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Techniques for Characterizing Cytomegalovirus-Encoded miRNAs.

Nicole L Diggins1, Lindsey B Crawford1, Hillary M Struthers1

  • 1Vaccine and Gene Therapy Institute, Oregon Health and Sciences University, Beaverton, OR, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 8, 2021
PubMed
Summary

This chapter details methods for studying microRNAs (miRNAs), small molecules that regulate gene expression. It focuses on techniques for detecting, expressing, inhibiting, and identifying targets of human cytomegalovirus-encoded miRNAs.

Keywords:
Human cytomegalovirus (HCMV)Locked nucleic acids (LNA)Luciferase assayNorthern BlotPhotoactivatable ribonucleoside-enhanced cross-linking and Immunoprecipitation (PAR CLIP)RISC-immunoprecipitation (RISC-IP)Stem-loop real-time PCRmicroRNA (miRNA)

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Area of Science:

  • Molecular Biology
  • Genetics
  • Bioinformatics

Background:

  • microRNAs (miRNAs) are small noncoding RNAs regulating gene expression posttranscriptionally.
  • miRNA binding to messenger RNA (mRNA) 3' untranslated regions impacts gene function.
  • The unique regulatory role of miRNAs necessitates advanced characterization techniques.

Purpose of the Study:

  • To review current techniques for detecting, expressing, and inhibiting miRNAs.
  • To discuss methods for identifying and validating miRNA targets.
  • To specifically address techniques relevant to human cytomegalovirus (HCMV)-encoded miRNAs.

Main Methods:

  • Review of established and emerging miRNA detection and quantification assays.
  • Description of methods for miRNA mimic and inhibitor design and delivery.
  • Overview of bioinformatic and experimental approaches for miRNA target prediction and validation.

Main Results:

  • The chapter provides a comprehensive overview of the miRNA research toolkit.
  • It highlights the specific challenges and methodologies applicable to viral miRNAs.
  • Key techniques for analyzing miRNA-mediated gene regulation are presented.

Conclusions:

  • A robust set of techniques exists for miRNA research.
  • Understanding HCMV-miRNA interactions requires specialized methods.
  • This chapter serves as a guide to miRNA characterization and target validation.