A Cross-Tissue Investigation of Molecular Targets and Physiological Functions of Nsun6 Using Knockout Mice
Abstract
:1. Introduction
2. Results
2.1. Nsun6 Gene Is Not Essential for the Development and Breeding of Mice
2.2. Identification of High-Confident Nsun6-Dependent m5C Sites across Tissues
2.3. Nsun6 Deficiency Induces Limited Transcriptome Changes across Tissues
2.4. Nsun6-KO Impairs the Formation of Antibody-Secreting Plasma Cells in the Spleen
2.5. Nsun6-Mediated m5C Modification Does Not Affect RNA Stability across Tissues
3. Discussion
4. Methods
4.1. Generation of the Nsun6-Knockout Mouse Strain
4.2. Total RNA Extraction
4.3. mRNA Capture
4.4. mRNA Library Construction
4.5. mRNA Bisulfite Sequencing
4.6. Western Blot
4.7. Detection of the m5C Modification
4.8. Nsun6-Dependent Site Identification
4.9. Mice Immunization
4.10. Flow Cytometry
4.11. ELISPOT Assay
4.12. RNA-seq Analysis
4.13. t-SNE Visualization
4.14. GO Enrichment Analysis
4.15. GSEA Analysis
4.16. Motif Analysis of m5C Sites
4.17. Data Availability
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Wang, W.; Huang, H.; Jiang, H.; Tian, C.; Tang, Y.; Gan, D.; Wen, X.; Song, Z.; He, Y.; Ou, X.; et al. A Cross-Tissue Investigation of Molecular Targets and Physiological Functions of Nsun6 Using Knockout Mice. Int. J. Mol. Sci. 2022, 23, 6584. https://doi.org/10.3390/ijms23126584
Wang W, Huang H, Jiang H, Tian C, Tang Y, Gan D, Wen X, Song Z, He Y, Ou X, et al. A Cross-Tissue Investigation of Molecular Targets and Physiological Functions of Nsun6 Using Knockout Mice. International Journal of Molecular Sciences. 2022; 23(12):6584. https://doi.org/10.3390/ijms23126584
Chicago/Turabian StyleWang, Wen, Hengjun Huang, Hao Jiang, Chi Tian, Yisen Tang, Diwen Gan, Xiaozhen Wen, Zhenyu Song, Yuhao He, Xijun Ou, and et al. 2022. "A Cross-Tissue Investigation of Molecular Targets and Physiological Functions of Nsun6 Using Knockout Mice" International Journal of Molecular Sciences 23, no. 12: 6584. https://doi.org/10.3390/ijms23126584