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Related Concept Videos

RNA Splicing01:32

RNA Splicing

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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...
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Alternative RNA Splicing02:18

Alternative RNA Splicing

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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
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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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Exon Recombination02:32

Exon Recombination

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The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon...
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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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Pre-mRNA Processing: RNA Splicing01:36

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Updated: Sep 6, 2025

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
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Engineered U1 snRNAs to modulate alternatively spliced exons.

Samuel T Hatch1, Aaron A Smargon2, Gene W Yeo1

  • 1Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA; Stem Cell Program, University of California San Diego, Sanford Consortium for Regenerative Medicine, La Jolla, USA; Institute for Genomic Medicine, University of California San Diego, La Jolla, CA, USA; Biomedical Sciences Graduate Program, University of California San Diego, La Jolla, CA, USA.

Methods (San Diego, Calif.)
|June 28, 2022
PubMed
Summary
This summary is machine-generated.

Engineered U1 small nuclear RNAs (snRNAs) offer a promising therapeutic approach for correcting errors in mRNA splicing. This study presents a user-friendly toolkit demonstrating robust modulation of pre-mRNA splicing for research and potential therapeutic applications.

Keywords:
ASOAlternative splicingAntisenseExonExonicFASGene therapyGuideIntronIntronicRNARNA targetingSMN2Sequence-specificSmall nuclearSplice siteSpliceosomeSplicingSplicing enhancerSplicing silencerTranscriptTranscriptionU snRNAU1 snRNAUridine-richmRNApre-mRNAsnRNAsnRNP

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Area of Science:

  • Molecular Biology
  • Genetics
  • RNA Biology

Background:

  • Alternative splicing generates transcriptomic diversity, but errors lead to diseases like spinal muscular atrophy.
  • Small nuclear RNAs (snRNAs), particularly U1 snRNAs, are being explored as therapeutics for splicing defects.
  • Previous in vivo studies show U1 snRNA efficacy, warranting further preclinical development.

Purpose of the Study:

  • To develop and demonstrate a user-friendly methodological toolkit for targeted perturbation of alternative splicing using engineered U1 snRNAs.
  • To validate the utility of engineered U1 snRNAs in modulating endogenous pre-mRNA splicing in different cellular contexts.

Main Methods:

  • Development of a methodological toolkit for sequence-specific targeting of pre-mRNA.
  • Utilizing engineered U1 snRNAs to modulate alternative splicing.
  • Testing the toolkit on SMN2 exon 7 and FAS exon 6 splicing contexts.

Main Results:

  • Robust modulation of endogenous pre-mRNA transcripts was observed.
  • Demonstrated successful application of engineered U1 snRNA for both exon inclusion and exclusion.
  • Validated the utility and applicability of the engineered U1 snRNA toolkit across different splicing contexts.

Conclusions:

  • Engineered U1 snRNAs provide a versatile tool for investigating and potentially treating splicing-related disorders.
  • The presented toolkit enhances the accessibility and usability of U1 snRNA technology for the research community.
  • Further advancement of U1 snRNA technology is justified for therapeutic and research applications in splicing modulation.