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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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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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Nonsense-mediated mRNA Decay02:27

Nonsense-mediated mRNA Decay

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The Upf proteins that carry out nonsense-mediated decay (NMD) are found in all eukaryotic organisms, including humans. Each protein has an individual role, but they need to work in collaboration. Upf1 is an ATP-dependent RNA helicase that unwinds the RNA helix. Because Upf1 can unwind any RNA, Upf2 and Upf3 are required to help Upf1 discriminate between nonsense and normal mRNAs.
Usually, Upf3 binds to an Exon Junction Complex (EJC) at mRNA splice sites. If a ribosome fully translates the mRNA,...
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Regulation of Nuclear Protein Sorting01:45

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Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
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Related Experiment Video

Updated: Jan 14, 2026

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
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A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

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Small molecule splicing modulators that disrupt O-GlcNAc homeostasis.

Steven S Cheng1, Alison C Mody1, Amedeo Vetere2

  • 1Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.

Nature Communications
|January 12, 2026
PubMed
Summary

Drug repurposing screens identified kinase inhibitors that disrupt O-GlcNAc homeostasis by modulating splicing. These compounds downregulate O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) independently of their known targets.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • O-Linked N-acetylglucosamine (O-GlcNAc) is a crucial post-translational modification regulated by OGT and OGA.
  • Dysregulation of O-GlcNAc is implicated in various human diseases, driving therapeutic interest.
  • Drug repurposing offers a strategy to identify compounds that modulate O-GlcNAc pathways.

Purpose of the Study:

  • To identify clinically relevant compounds that can therapeutically modulate O-GlcNAc homeostasis.
  • To investigate the intersection of drug repurposing candidates with O-GlcNAc cycling enzymes.
  • To uncover novel mechanisms for disrupting O-GlcNAc homeostasis.

Main Methods:

  • Conducted three parallel drug repurposing screens in cells and in vitro targeting O-GlcNAc cycling enzymes.
  • Utilized kinase inhibitors GSK690693 and Y-33075 to assess their impact on O-GlcNAc homeostasis.
  • Evaluated a panel of splicing modulators for their effects on OGT and OGA expression.

Main Results:

  • GSK690693 and Y-33075 were identified as splicing modulators that disrupt O-GlcNAc homeostasis by downregulating OGT and OGA.
  • These effects were independent of the inhibitors' known targets (AKT and ROCK) and distinct from direct OGT/OGA inhibitors.
  • Additional splicing modulators (OTS964, indisulam, GNF2133) were found to downregulate OGT and OGA with unique splicing profiles.

Conclusions:

  • Kinase inhibitors can act as splicing modulators to disrupt O-GlcNAc homeostasis.
  • Drug repurposing screens revealed novel chemotypes targeting O-GlcNAc cycling enzymes through splicing modulation.
  • These findings provide new therapeutic strategies for diseases associated with O-GlcNAc dysregulation.