Related Experiment Video

Updated: Jun 23, 2025

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
06:48

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells

Published on: June 16, 2022

2.0K

MicroRNA biogenesis is broadly disrupted by inhibition of the splicing factor SF3B1

Angela Downie Ruiz Velasco1, Aimee L Parsons1, Matthew C Heatley2

  • 1School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, UK.

Nucleic Acids Research
|June 17, 2024
PubMed

Insights

Splicing inhibition significantly reduced the transcription and production of miR-122, a microRNA (miRNA) encoded by a long non-coding RNA. This study highlights the crucial role of splicing in miRNA biogenesis.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNA (miRNA) biogenesis is initiated by the Microprocessor complex's cleavage of primary miRNA (pri-miRNA).
  • The influence of cotranscriptional splicing on miRNA production is established for intronic miRNAs within protein-coding genes, but its role in long non-coding RNA (lncRNA)-hosted miRNAs remains unclear.

Purpose of the Study:

  • To investigate the impact of splicing on the biogenesis of miR-122, a critical liver-specific miRNA hosted in a lncRNA.
  • To elucidate the role of the SF3B1 protein in miRNA production.

Main Methods:

  • Utilized pladienolide B (PlaB), an SF3B1 inhibitor, to impede splicing.
  • Employed SLAMseq technology to quantify global miRNA synthesis rates following short-term splicing inhibition.
  • Compared the effects of splicing inhibition on endogenous and plasmid-encoded pri-miR-122 transcription.

Main Results:

  • Splicing inhibition by PlaB markedly reduced endogenous pri-miR-122 transcription and subsequent mature miR-122 production.
  • SLAMseq analysis revealed an overall decrease in mature miRNA biogenesis upon PlaB treatment.
  • Both exonic and intronic miRNAs were similarly affected by splicing inhibition.

Conclusions:

  • Splicing plays a significant role in promoting the biogenesis of lncRNA-hosted miRNAs like miR-122.
  • SF3B1 is essential for the global biogenesis of miRNAs.
  • This study reveals novel biological insights into the interplay between splicing and transcription in miRNA production.

More Related Videos

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

2.7K
Using the E1A Minigene Tool to Study mRNA Splicing Changes10:25

Using the E1A Minigene Tool to Study mRNA Splicing Changes

Published on: April 22, 2021

4.9K

Related Concept Videos

MicroRNAs01:22

MicroRNAs

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...
3.0K
RNA Interference01:23

RNA Interference

RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
26.0K
Experimental RNAi02:15

Experimental RNAi

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...
6.1K
RNA Splicing01:32

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
56.3K
siRNA - Small Interfering RNAs02:30

siRNA - Small Interfering RNAs

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.
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...
16.7K
Chromatin Structure Regulates pre-mRNA Processing02:41

Chromatin Structure Regulates pre-mRNA Processing

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...
7.0K