miR-541-3p Promoted Porcine Reproductive and Respiratory Syndrome Virus 2 (PRRSV-2) Replication by Targeting Interferon Regulatory Factor 7
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Virus
2.2. Antibodies and Reagents
2.3. Plasmids
2.4. RNA Quantification
2.5. Dual Luciferase Reporter Assay
2.6. RNA Interference Assay
2.7. Virus Titration
2.8. Statistical Analysis
3. Results
3.1. miR-541-3p Facilitated the Replication of PRRSV-2 in MARC-145 Cells
3.2. PRRSV-2 Infection Up-Regulated the Expression of miR-541-3p in MARC-145 Cells
3.3. miR-541-3p Negatively Regulated Poly(I:C)-Induced Transcription of Type I Interferon
3.4. Interferon Regulatory Factor (IRF) 7 Was a Target Gene of miR-541-3p
3.5. PRRSV-2 Infection Down-Regulated the Expression of IRF7 and the Over-Expression of IRF7 Could Inhibit the Replication of PRRSV-2 in MARC-145 Cells
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Primers | Sequence (5′-3′) |
---|---|
MAVS-For | CTGCCTCACAGCAAGAGACCA |
MAVS-Rev | GTAGACACAGGCCACTTCGTC |
IRAK2-For | AGGCACGGGAAGCCATTCGT |
IRAK2-Rev | AGCCCAGCACAGGTAAGACA |
IRAK3-For | TTGGTCCTGGGCACAGAAA |
IRAK3-Rev | AATAGCTCGACGATGCCCCA |
IRAK4-For | TTGCTCAGGGTGCAGCTAAT |
IRAK4-Rev | GTGCAAGGCCAAAGTCAGAT |
TRAF3-For | CTCACAAATGCAGCGTCCAG |
TRAF3-Rev | GCTCCACTCCTTCAGCAGGT |
IRF7-For | GCAACGTGAGGGTGTGTCTT |
IRF7-Rev | GCTCCATGAGCAAGCACTCAA |
IFNAR1-For | TCGTTTACACCATTTCGCAAAGCTCAAA |
IFNAR1-Rev | ACGATCCAAAGCCCACATGACACTATCT |
IFNAR2-For | TGTCATTGAAGAGCAGTCAGAGGGGATT |
IFNAR2-Rev | GTTGAAGGAGGGTGCATTTTAAGGGAGA |
GAPDH-For | TGACAACAGCCTCAAGATCG |
GAPDH-Rev | GTCTTCTGGGTGGCAGTGAT |
ORF7-For | AAACCAGTCCAGAGGCAAGG |
ORF7-Rev | GCAAACTAAACTCCACAGTGTAA |
IFN-β-For | CTAGCACTGGCTGGAATGAGACT |
IFN-β-Rev | GGCCTTCAGGTAATGCAGAATC |
Flag-IRF7-For | CCCAAGCTTATGGCCTTGGCTCCTGAGAG |
Flag-IRF7-Ror | CGCGGATCCCTAGACGGGCTGCTCCAGCTC |
Top strand oligo for miR-541-3p | GGATCCTGGAGGCTTGCTGAAGGCTGTATGCTGTGGTGGGCACAGAATCTGGACTGTTTTGGCCACTGACTGAC |
Bottom strand oligo for miR-541-3p | CAAAACCGGTGACTGACTGTCAGGTCTGACACGGGTGGTGTCCTGTGTTCCGGACAATGATCGTGAGTGTACCTTGTTTACCGGGTCTAGA |
pcDNA6.2-miR-541-3p-For | GGATCCTGGAGGCTTGCTGAA |
pcDNA6.2-miR-541-3p-Rev | AGATCTGGGCCATTTGTTCCATGT |
Top strand oligo for mut-miR-541-3p | GGATCCTGGAGGCTTGCTGAAGGCTGTATGCTGTCTCTTACACAGAATCTGGACTGTTTTGGCCACTGACTGAC |
Bottom strand oligo for mut-miR-541-3p | CAAAACCGGTGACTGACTGTCAGGTCTGACACATTCTCTGTCCTGTGTTCCGGACAATGATCGTGAGTGTACCTTGTTTACCGGGTCTAGA |
IRF7-3′UTR For | TCGAACTCCAGAAAGCTGGAGCAGCCCACCTAGAGCTGGCCGC |
IRF7-3′UTR Rev | GGCCGCGGCCAGCTCTAGGTGGGCTGCTCCAGCTTTCTGGAGT |
Mut-IRF7-3′UTR | TCGAACTCCAGAAAGCTGGAGCAAGATCGTTAGAGCTGGCCGC |
Mut-IRF7-3′UTR | GGCCGCGGCCAGCTCTAACGATCTTGCTCCAGCTTTCTGGAGT |
siIRF7-sense | CCAUCUUUGACUUCAGAGUTT |
siIRF7-antisense | ACUCUGAAGUCAAAGAUGGTT-3 |
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Shi, X.; Yang, Y.; Zhang, X.; Chang, X.; Chen, J.; Wang, C.; Wang, A.; Wang, J.; Qin, J.; Ye, X.; et al. miR-541-3p Promoted Porcine Reproductive and Respiratory Syndrome Virus 2 (PRRSV-2) Replication by Targeting Interferon Regulatory Factor 7. Viruses 2022, 14, 126. https://doi.org/10.3390/v14010126
Shi X, Yang Y, Zhang X, Chang X, Chen J, Wang C, Wang A, Wang J, Qin J, Ye X, et al. miR-541-3p Promoted Porcine Reproductive and Respiratory Syndrome Virus 2 (PRRSV-2) Replication by Targeting Interferon Regulatory Factor 7. Viruses. 2022; 14(1):126. https://doi.org/10.3390/v14010126
Chicago/Turabian StyleShi, Xibao, Yuanhao Yang, Xiaozhuan Zhang, Xiaobo Chang, Jing Chen, Chao Wang, Aiping Wang, Jianhua Wang, Jianru Qin, Xianlong Ye, and et al. 2022. "miR-541-3p Promoted Porcine Reproductive and Respiratory Syndrome Virus 2 (PRRSV-2) Replication by Targeting Interferon Regulatory Factor 7" Viruses 14, no. 1: 126. https://doi.org/10.3390/v14010126