Human Cancer-Associated Mutations of SF3B1 Lead to a Splicing Modification of Its Own RNA
Abstract
:1. Introduction
2. Results and Discussion
2.1. Expression of the Main SF3B1 Mutations in MDS Patients and in Myeloid Cell Lines Drives the Formation of SF3B1ins Transcript
2.2. SF3B1ins and SF3B1K700Eins Proteins Are Defective for Splicing
2.3. Study of Splicing Defects due to Expression of SF3B1ins or Disease-Related SF3B1 Mutations in Schizosaccharomyces pombe
2.4. Use of Splice-Switching Oligonucleotides to Modulate the Production of SF3B1ins Transcript
3. Materials and Methods
3.1. Patient Samples
3.2. Next-Generation Sequencing
3.3. Cloning and Site-Directed Mutagenesis
3.4. Cell Culture and Transfection
3.5. Generation of SF3B1 and shSF3B1 Inducible Cell Lines
3.6. RNA Extraction and RT-PCR
3.7. Western Blot Analysis
3.8. Yeast Growth and Molecular Biology
3.9. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Bergot, T.; Lippert, E.; Douet-Guilbert, N.; Commet, S.; Corcos, L.; Bernard, D.G. Human Cancer-Associated Mutations of SF3B1 Lead to a Splicing Modification of Its Own RNA. Cancers 2020, 12, 652. https://doi.org/10.3390/cancers12030652
Bergot T, Lippert E, Douet-Guilbert N, Commet S, Corcos L, Bernard DG. Human Cancer-Associated Mutations of SF3B1 Lead to a Splicing Modification of Its Own RNA. Cancers. 2020; 12(3):652. https://doi.org/10.3390/cancers12030652
Chicago/Turabian StyleBergot, Tiffany, Eric Lippert, Nathalie Douet-Guilbert, Séverine Commet, Laurent Corcos, and Delphine G. Bernard. 2020. "Human Cancer-Associated Mutations of SF3B1 Lead to a Splicing Modification of Its Own RNA" Cancers 12, no. 3: 652. https://doi.org/10.3390/cancers12030652