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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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siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

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Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements.
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Updated: Sep 15, 2025

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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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.

Biorxiv : the Preprint Server for Biology
|July 14, 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 in precursor miR-20a. These features control enzymatic processing and are affected by disease-related genetic variations.

Keywords:
DicerNMR spectroscopyRNA structureSAXSmicroRNA

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Genetics

Background:

  • MicroRNAs (miRNAs) are crucial regulators of gene expression, and their precise processing is essential for cellular function.
  • The molecular mechanisms governing miRNA biogenesis, particularly the processing of precursor miRNAs (pre-miRNAs), are not fully elucidated.
  • Pre-miR-20a, a component of the oncomiR-1 cluster, is an oncogenic miRNA implicated in disease.

Purpose of the Study:

  • To determine the solution structure of pre-miR-20a to understand its processing regulation.
  • To identify structural features of pre-miR-20a that influence its enzymatic maturation.
  • To investigate the impact of a disease-related single-nucleotide polymorphism on pre-miR-20a processing.

Main Methods:

  • Nuclear magnetic resonance (NMR) spectroscopy to determine the solution structure of pre-miR-20a.
  • Small-angle X-ray scattering (SAXS) to complement structural analysis.
  • Analysis of pre-miR-20a structure and its interaction with processing enzymes (Dicer-TRBP).

Main Results:

  • The solution structure of pre-miR-20a revealed a flexible apical loop and a single-nucleotide bulge near the Dicer-TRBP cleavage site.
  • Alternative conformations in the apical loop were identified as key self-regulatory elements for Dicer-TRBP processing.
  • A single-nucleotide bulge at the -5 position is critical for efficient pre-miR-20a processing.
  • A disease-related single-nucleotide polymorphism (SNP) disrupting the structure near the cleavage site led to reduced processing efficiency.

Conclusions:

  • Transient RNA conformations within 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 basis for understanding oncomiR regulation and disease-associated genetic variations.