Identification of Differential Circular RNA Expression Profiles and Functional Networks in Human Macrophages Induced by Virulent and Avirulent Mycobacterium tuberculosis Strains
Abstract
:1. Introduction
2. Results
2.1. CircRNA Profiling in All THP-1 Cells
2.2. Analysis of Differentially Expressed circRNAs
2.3. KEGG and Reactome Pathway Analysis of Differentially Expressed circRNA Parental Genes
2.4. Detection of Differentially Expressed circRNAs
2.5. Construction of M.tb Infection-Related circRNA-miRNA-mRNA Competing Endogenous Interaction Network
3. Discussion
3.1. CircRNAs Expression in Macrophages
3.2. Differential circRNAs Associated with Virulent and Avirulent M.tb
3.3. Predicted Interaction Network of Novel Identified circRNAs in M.tb-Infected Cells
3.4. Limitation
4. Materials and Methods
4.1. Bacterial Strains and Cell Culture
4.2. Cell Infection with Bacteria
4.3. CircRNA Sequencing and Differential Expression Profile Analysis
4.4. RNase R Digestion Analyses
4.5. Reverse Transcription Polymerase Chain Reaction (RT-PCR)
4.6. Quantitative Real-Time PCR (qPCR)
4.7. Nucleocytoplasmic Isolation
4.8. KEGG and Reactome Enrichment Analyses
4.9. Prediction of miRNA-circRNA and circRNA-miRNA-circRNA ceRNA Network
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer Names | Sequences (5′–3′) | Products (bp) |
---|---|---|
hsa-β-actin | F: CATGTACGTTGCTATCCAGGC R: CTCCTTAATGTCACGCACGAT | 250 |
hsa-NEAT1 | F: AAACGCTGGGAGGGTACAAG R: ATGCCCAAACTAGACCTGCC | 71 |
hsa-GAPDH | F: AATGGGCAGCCGTTAGGAAA R: GCCCAATACGACCAAATCAGAG | 166 |
hsa-circCHSY1- Divergent | F: AAGGTGTGTCCGGAGGTTTG R: TGGCACTACTGGAATTGGTACA | 134 |
hsa-circCHSY1- Convergent | F: TGCACGACCACTACTTGGAC R: GCCTGTCTGCCCAAGAAAGA | 134 |
hsa-circSOD2- Divergent | F: AATGTAATCAACTGGGAGAATG R: GGCTGTAACATCTCTCAGCATA | 87 |
hsa-circSOD2- Convergent | F: CTGGAAGCCATCAAACGTGAC R: AACCTGAGCCTTGGACACC | 88 |
hsa-circTNFRSF21- Divergent | F: TACTGCAATGGCCATGCTTG R: TTCCTGCTGGACACTTGTCA | 140 |
hsa-circTNFRSF21- Convergent | F: CTGCCTTGACTGACCGAGAA R: ACACTGCTTACACCGCACAT | 143 |
hsa-circDHTKD1- Divergent | F: CGTGGTCGTTTGTTTCTCCA R: TGGCAGCTTTATGACCATGC | 107 |
hsa-circDHTKD1- Convergent | F: TGCTTACAGGTCCATGGTGA R: ATGCACACTCCCACCAATTC | 105 |
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Zhu, Y.; Kong, D.; Wang, Z.; Li, T.; Tang, T.; Peng, Y.; Hu, C.; Chao, J.; Chen, H.; Chen, Y.; et al. Identification of Differential Circular RNA Expression Profiles and Functional Networks in Human Macrophages Induced by Virulent and Avirulent Mycobacterium tuberculosis Strains. Int. J. Mol. Sci. 2023, 24, 17561. https://doi.org/10.3390/ijms242417561
Zhu Y, Kong D, Wang Z, Li T, Tang T, Peng Y, Hu C, Chao J, Chen H, Chen Y, et al. Identification of Differential Circular RNA Expression Profiles and Functional Networks in Human Macrophages Induced by Virulent and Avirulent Mycobacterium tuberculosis Strains. International Journal of Molecular Sciences. 2023; 24(24):17561. https://doi.org/10.3390/ijms242417561
Chicago/Turabian StyleZhu, Yifan, Delai Kong, Zijian Wang, Ting Li, Tian Tang, Yongchong Peng, Changmin Hu, Jin Chao, Huanchun Chen, Yingyu Chen, and et al. 2023. "Identification of Differential Circular RNA Expression Profiles and Functional Networks in Human Macrophages Induced by Virulent and Avirulent Mycobacterium tuberculosis Strains" International Journal of Molecular Sciences 24, no. 24: 17561. https://doi.org/10.3390/ijms242417561