siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Virus
2.2. Viral Infections and Plaque Assays
2.3. Quantitative Real Time-PCR
2.4. Western Blot
2.5. Small Interferring RNA Design
2.6. Small Interfering RNA Silencing
2.7. Cell Viability Assay
3. Results
3.1. Alignment of the Human Respiratory Syncytial Virus Genome with Innate Immune Genes and Selection of Bispecific Small Interfering RNAs
3.2. Small Interfering RNAs Targeting F or N Inhibit Virus Growth
3.3. Silencing N, but Not F, Leads to a Generalized Inhibition of Human Respiratory Syncytial Virus RNA and Proteins
3.4. Targeting Human Respiratory Syncytial Virus Nucleoprotein with Small Interfering RNAs Drastically Reduces the Cellular Innate Immune Response While Human Respiratory Syncytial Virus Fusion Protein Silencing Has Only a Moderate Effect
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Name a | Sense Sequence (5′-3′) b | Antisense Sequence (5′-3′) b |
---|---|---|
TRIM25 | CAACAAGAAUACACGGAAAtt | UUUCCGUGUAUUCUUGUUGta |
HRSV-F | GCAAAGUGUUAGACCUCAAtt | UUGAGGUCUAACACUUUGCtg |
TRIM25/HRSV-F | CCAUAGACCUCAAAAACGAtt | UCGUUUUUGAGGUCUAUGGtg |
RIG-I | CAAGAAGAGUACCACUUAAtt | UUAAGUGGUACUCUUCUUGta |
HRSV-N (ALN-RSV01) | GGCUCUUAGCAAAGUCAAGtt | CUUGACUUUGCUAAGAGCCtt |
RIG-I/HRSV-N | GGAAAUGGAGCAAGUUGUUtt | AACAACUUGCUCCAUUUCCtc |
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Martín-Vicente, M.; Resino, S.; Martínez, I. siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication. Biomolecules 2019, 9, 165. https://doi.org/10.3390/biom9050165
Martín-Vicente M, Resino S, Martínez I. siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication. Biomolecules. 2019; 9(5):165. https://doi.org/10.3390/biom9050165
Chicago/Turabian StyleMartín-Vicente, María, Salvador Resino, and Isidoro Martínez. 2019. "siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication" Biomolecules 9, no. 5: 165. https://doi.org/10.3390/biom9050165