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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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pre-mRNA Processing02:01

pre-mRNA Processing

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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
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Pre-mRNA Processing: Modification of pre-mRNA Ends01:35

Pre-mRNA Processing: Modification of pre-mRNA Ends

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In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps...
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Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

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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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Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

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In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
The chromatin structure, especially...
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Experimental RNAi02:15

Experimental RNAi

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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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Structural features within precursor microRNA-20a regulate Dicer-TRBP processing.

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Updated: Sep 17, 2025

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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Structural Features Within Precursor microRNA-20a Regulate Dicer-TRBP Processing.

Yaping Liu1, Cade T Harkner2, Megan N Westwood1

  • 1Biophysics Program, University of Michigan, 930 N. University Avenue, Ann Arbor, MI 48109, USA.

Journal of Molecular Biology
|July 3, 2025
PubMed
Summary
This summary is machine-generated.

MicroRNA (miRNA) processing is regulated by structural elements like the apical loop and a specific nucleotide bulge. These features control enzymatic processing and are crucial for proper gene expression regulation.

Keywords:
DicerNMR spectroscopyRNA structureSAXSmicroRNA

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Genetics

Background:

  • MicroRNAs (miRNAs) are small non-coding RNAs regulating gene expression post-transcriptionally.
  • Precise control of miRNA biogenesis is essential for maintaining cellular homeostasis.
  • The molecular mechanisms governing miRNA precursor processing remain incompletely understood.

Purpose of the Study:

  • To elucidate the structural determinants of pre-miR-20a processing.
  • To understand how structural features regulate the enzymatic maturation of an oncogenic miRNA.

Main Methods:

  • Solution structure determination of pre-miR-20a using Nuclear Magnetic Resonance (NMR) spectroscopy.
  • Small Angle X-ray Scattering (SAXS) analysis.
  • Assessment of pre-miR-20a processing efficiency.

Main Results:

  • Identified a flexible apical loop and a single-nucleotide bulge near the Dicer-TRBP cleavage site in pre-miR-20a.
  • Demonstrated that alternative apical loop conformations self-regulate Dicer-TRBP processing.
  • Showed that a bulge at the -5 position is critical for efficient pre-miR-20a processing.
  • Found that a disease-related single-nucleotide polymorphism impairs pre-miR-20a processing due to structural disruption.

Conclusions:

  • Transient RNA conformations in pre-miR-20a's apical loop self-regulate its maturation.
  • Specific structural features, including a nucleotide bulge, are critical for efficient miRNA processing.
  • Structural insights into pre-miR-20a provide a foundation for understanding oncomiR-1 cluster regulation and disease-related mutations.