The Fusion of CLEC12A and MIR223HG Arises from a trans-Splicing Event in Normal and Transformed Human Cells
Abstract
:1. Introduction
2. Results
2.1. CLEC12A-MIR223HG Is Expressed in CML Patients and Healthy Controls
2.2. CLEC12A-MIR223HG Is Expressed in Normal Human Tissues and Leukemic Cell Lines
2.3. CLEC12A-MIR223HG Exhibits a Typical Fusion Transcript Architecture
2.4. CRISPR-Mediated Transcriptional Activation of CLEC12A Increases CLEC12A-MIR223HG Expression
2.5. CLEC12A-MIR223HG Encodes a Chimeric Protein That Is Distinct from CLEC12A
2.6. Increased CLEC12A-MIR223HG Expression following Cell Differentiation or Chemotherapy
3. Discussion
4. Materials and Methods
4.1. Clinical Samples and Bioinformatics Analysis
4.2. Cell Lines and Culture
4.3. Expression Vectors, Lentivirus Production, and Transduction
4.4. RNA Isolation and RT-PCR
4.5. Western Blotting
4.6. Immunofluorescence
4.7. Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS) Analysis
4.8. CRISPR-SAM-Mediated Activation of CLEC12A and Analysis of Chromosomal Translocations in HEK293 Cells
4.9. U-937 Monocyte Differentiation Assay
4.10. Cell Apoptosis with Annexin V Staining
4.11. Cell Viability Assay
4.12. Colony Formation-Methocult Assay
4.13. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Dhungel, B.P.; Monteuuis, G.; Giardina, C.; Tabar, M.S.; Feng, Y.; Metierre, C.; Ho, S.; Nagarajah, R.; Fontaine, A.R.M.; Shah, J.S.; et al. The Fusion of CLEC12A and MIR223HG Arises from a trans-Splicing Event in Normal and Transformed Human Cells. Int. J. Mol. Sci. 2021, 22, 12178. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222212178
Dhungel BP, Monteuuis G, Giardina C, Tabar MS, Feng Y, Metierre C, Ho S, Nagarajah R, Fontaine ARM, Shah JS, et al. The Fusion of CLEC12A and MIR223HG Arises from a trans-Splicing Event in Normal and Transformed Human Cells. International Journal of Molecular Sciences. 2021; 22(22):12178. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222212178
Chicago/Turabian StyleDhungel, Bijay P., Geoffray Monteuuis, Caroline Giardina, Mehdi S. Tabar, Yue Feng, Cynthia Metierre, Sarah Ho, Rajini Nagarajah, Angela R. M. Fontaine, Jaynish S. Shah, and et al. 2021. "The Fusion of CLEC12A and MIR223HG Arises from a trans-Splicing Event in Normal and Transformed Human Cells" International Journal of Molecular Sciences 22, no. 22: 12178. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms222212178