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Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
Published on: April 26, 2017
Specific G-quadruplex ligands modulate the alternative splicing of Bcl-X
Carika Weldon1, Justine G Dacanay1, Vijay Gokhale2
1Leicester Institute of Structural & Chemical Biology and Department of Molecular & Cell Biology, University of Leicester, Leicester LE1 7RH, UK.
RNA G-quadruplexes (G4s) in pre-mRNA influence alternative splicing. Specific G4 ligands modulate Bcl-X pre-mRNA splicing, with one ligand inducing apoptosis by altering splice site selection.
Area of Science:
- Molecular Biology
- RNA Biology
- Genetics
Background:
- RNA G-quadruplexes (G4s) are prevalent in mammalian introns near splice sites.
- Formation and functional impact of G4s in pre-mRNA remain challenging to study.
- Previous work identified two G4s in Bcl-X pre-mRNA near alternative 5' splice sites (5'SS).
Purpose of the Study:
- To investigate the effect of G4 ligands on Bcl-X pre-mRNA splicing.
- To determine the specificity and mechanism of action of G4 ligands on alternative splicing.
- To explore the role of G4s in regulating the expression of Bcl-X isoforms.
Main Methods:
- Utilized specific G4 ligands to stabilize G4 structures in Bcl-X pre-mRNA.
- Analyzed changes in Bcl-X pre-mRNA splicing patterns in response to G4 ligands.
- Investigated the independent and combined effects of ligands on alternative 5'SS usage.
Main Results:
- A subset of G4 ligands significantly altered Bcl-X pre-mRNA splicing.
- Ligand activity and specificity were highly dependent on their chemical structures.
- The ellipticine GQC-05 ligand selectively modulated the two alternative 5'SS, favoring the pro-apoptotic isoform.
Conclusions:
- G4 structures in pre-mRNA can be targeted by small molecules to control alternative splicing.
- G4 ligands demonstrate specificity in modulating splice site selection, impacting isoform expression.
- The observed effects of GQC-05 suggest a mechanism for G4-mediated apoptosis induction through altered Bcl-X splicing.
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