A Short 5′triphosphate RNA nCoV-L Induces a Broad-Spectrum Antiviral Response by Activating RIG-I
Abstract
:1. Introduction
2. Materials and Methods
2.1. Biosafety and Ethics Approval
2.2. Cell Culture
2.3. Viruses
2.4. Mice
2.5. In Vitro Transcription and Gel Analysis
- 5′-GGUUUAAUACCUAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
- UCCCUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUAGGUAUUAAA
- CC-3′ (100 nt).
2.6. Transfections
2.7. Luciferase Assays
2.8. Protein Extraction and Western Blot
2.9. RNA Extraction
2.10. Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR)
2.11. In Vitro Virus Infection
2.12. In Vivo Virus Infection
2.13. Histopathological Examination
2.14. ELISA
2.15. Statistical Analyses
3. Results
3.1. nCoV-L Induce Interferon Response via the RIG-I Receptor
3.2. In Vitro Antiviral Effects of nCoV-L against RNA Viruses EV-71, CV-B5, PV-1 and IAV
3.3. nCoV-L Protected Suckling Mice from Lethal EV71 Virus Infection
3.4. Prophylactic Administration of nCoV-L Protects Mice from Lethal SARS-CoV-2 Virus Infection
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A
Gene | Wild-Type Sequence | Post-Editing Sequence | Mutation Type |
---|---|---|---|
TLR3 | CAACTTTCTTGGGACTA----AAGTGGACAAATCTCAC | CAACTTTCTTGGGACTAATTTAAGTGGACAAATCTCAC | frame shift |
TLR7 | AACTCTCTTCAGCATGTGCCCCCAAGATGGTTT | AACTCTCTT-------------CAAGATGGTTT | frame shift |
TLR8 | ATTTAGTGGGAGAAATAGC--CTCTGGGGCATTTTT | ATTTAGTGGGAGAAATAGCGCCTCTGGGGCATTTTT | frame shift |
RIG-I | AAAAAATTCCCACAAGGACAAAAGGGGAAAG | AAAAAATTCCCACAAGGACAA--GGGGAAAG | frame shift |
MDA5 | CCCCGGAGCCAGAACTCCAGCTCAGGCCTTACCAA | CCCCGGAGCCAGAACTCC----CAGGCCTTACCAA | frame shift |
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Gene | sgRNA |
---|---|
TLR3 | CAACTTTCTTGGGACTAAAG |
TLR7 | TTCAGCATGTGCCCCCAAGA |
TLR8 | TTAGTGGGAGAAATAGCCTC |
MDA5 | GCTCAGGCCTTACCAAATGG |
RIG-I | AATTCCCACAAGGACAAAAG |
Gene | Direction | Primer Sequence (5′-3′) |
---|---|---|
EV71/C2-2 | Forward | TAG TTT CTT CAG CAG GGC GG |
Reverse | ACC ATT GGT AAG CAC TCG CA | |
CV-B5/JS417 | Forward | GGT GTC CGT GTT TCC TTT TAT TCC TAC |
Reverse | CAA GTA GAT AAT AGC TCT GTT TGT CAC CG | |
PV-1/Sabin | Forward | TGA TCA CAA CCC GAC CAA GG |
Reverse | TAA TCC ACT CCA GGG CCG TA | |
IAV/H9N2 | Forward | CCG GAA TTT CTG GAG AGG CG |
Reverse | TAC ACA AGC AGG CAA GCA GG | |
IFN-β | Forward | GCT TGG ATT CCT ACA AAG AAG CA |
Reverse | ATA GAT GGT CAA TGC GGC GTC |
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Song, Z.; Wang, Q.; Bian, L.; An, C.; Cui, B.; Mao, Q.; Wu, X.; He, Q.; Bai, Y.; Liu, J.; et al. A Short 5′triphosphate RNA nCoV-L Induces a Broad-Spectrum Antiviral Response by Activating RIG-I. Viruses 2022, 14, 2451. https://doi.org/10.3390/v14112451
Song Z, Wang Q, Bian L, An C, Cui B, Mao Q, Wu X, He Q, Bai Y, Liu J, et al. A Short 5′triphosphate RNA nCoV-L Induces a Broad-Spectrum Antiviral Response by Activating RIG-I. Viruses. 2022; 14(11):2451. https://doi.org/10.3390/v14112451
Chicago/Turabian StyleSong, Ziyang, Qian Wang, Lianlian Bian, Chaoqiang An, Bopei Cui, Qunying Mao, Xing Wu, Qian He, Yu Bai, Jianyang Liu, and et al. 2022. "A Short 5′triphosphate RNA nCoV-L Induces a Broad-Spectrum Antiviral Response by Activating RIG-I" Viruses 14, no. 11: 2451. https://doi.org/10.3390/v14112451