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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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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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Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
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The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
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Gene expression in prokaryotes is governed by constitutive and regulated systems, allowing cells to balance the production of essential proteins with adaptive responses to environmental changes.Constitutive Gene ExpressionConstitutive, or housekeeping, genes are continuously expressed as they encode proteins vital for fundamental cellular processes. These include enzymes for glycolysis, ribosomal components for protein synthesis, and proteins involved in DNA replication. Their constant...
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MicroRNA-based Regulation of Picornavirus Tropism
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Characterization of MicroRNAs Regulating FOXO Expression.

Doug Hanniford1,2, Eva Hernando3,4

  • 1Department of Pathology, New York University Langone Health, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 11, 2018
PubMed
Summary

This study details methods to confirm if microRNAs regulate gene expression, specifically testing if miR-182 targets FOXO3. These techniques aid in understanding microRNA-mediated gene regulation and identifying novel targets.

Keywords:
3′UTR reporter assayFOXOMicroRNAMicroRNA target identification

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Area of Science:

  • Molecular Biology
  • Genetics
  • Gene Regulation

Background:

  • MicroRNAs (miRNAs) are key post-transcriptional regulators influencing numerous genes.
  • The FOXO family of transcription factors is known to be regulated by miRNAs.
  • Understanding specific miRNA-target interactions is crucial for deciphering gene regulatory networks.

Purpose of the Study:

  • To present reproducible methods for validating miRNA-target interactions.
  • To demonstrate the direct targeting of the FOXO3 gene product by a candidate microRNA, miR-182.
  • To offer guidance on selecting candidate miRNAs and employing unbiased methods for miRNA-target identification.

Main Methods:

  • Utilizing 3' Untranslated Region (UTR) reporter assays to assess miRNA binding and repression.
  • Performing Western blotting to quantify protein levels following miRNA manipulation.
  • Implementing candidate miRNA selection strategies and unbiased miRNA-target identification approaches.

Main Results:

  • Established protocols for validating miRNA-mediated regulation of gene expression.
  • Provided evidence for direct targeting of FOXO3 by miR-182.
  • Outlined a framework for miRNA-target discovery.

Conclusions:

  • The described methods are generalizable for testing miRNA-target interactions.
  • miR-182 directly targets the FOXO3 gene product.
  • The study contributes valuable tools and insights for miRNA research.