Identification of Regulatory Host Genes Involved in Sigma Virus Replication Using RNAi Knockdown in Drosophila
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells, Flies, Virus Stock, and CO2 Assay
2.2. dsRNA Preparation
2.3. dsRNA Transfection, Virus Infection, and RNA Extraction
2.4. Gene Expression Assay Using RT-qPCR
2.5. dsRNA Microinjection
2.6. Statistical Analysis
3. Results
3.1. Viral Genome Replication Level Was Increased after the Knockdown of Immune Pathways in S2 Cells
3.2. Viral Gene Expression Increased after the Knockdown of Immune Pathways in S2 Cells
3.3. In Vivo Experiments Confirmed the Findings of in Vitro Experiments
3.4. TATA-Binding Protein May Facilitate Viral Replication by Enhancing the RNA Polymerase Activity
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene Name | dsRNA Synthesis Primers |
---|---|
domeless (dome) | F: TAATACGACTCACTATAGGG TAACGGCAAGAGCGC |
R: TAATACGACTCACTATAGGG AGGTTCTGGCCAGGT | |
PGRP-LC | F: TAATACGACTCACTATAGGGG GCGGTT TCCATACGG |
R: TAATACGACTCACTATAGGGG CCATTGCTGACGCTC | |
GFP | F: GCTCGGGAGATCTCCTGCCTTTGGGTGTGTCTGGG |
R: CTAGACTCGAGCGGCCAACGGATCCTTCGTAGCCC | |
TBP | F: AATTAACCCTCACTAAAGGGAT GGACCAAATGCTAAGCCC |
R: AATTAACCCTCACTAAAGGGTACTTTCTCGCTGCCAGTCT |
Gene | qPCR Check Primers | Gene | qPCR Check Primers |
---|---|---|---|
domeless (dome) | F: ACAACAGGCGTCTTCGGATT | SV-NP | F: TAACTCGGGTGTGACAGCTC |
R: ACCCTTCAGTTTTGCCATGGT | R: CTTCGTTCATCTTCCTGGGT | ||
PGRP-LC | F: CGCAAGGCCGTCACAGTTAC | SV-N | F: CACATGAGAAAATGCAAACAGCTT |
R: GGTTCAACGTCTTTCCGAAGAG | R: GAAAATGGAGCGAGGATCGA | ||
Diptericin (Dpt) | F: CTATTCATTGGACTGGCTTGTGCC | SV-P | F: TCAAACCCAGAGCCAGAGATAGTAT |
R: TGGAACTGGCGACGCACTCT | R: CGCTTTTATCTGACGCTCAGGTA | ||
TEPS | F: AACTCCGCAAACACCAAGTTGG | SV-X | F: TGGCCCCAATATTTCCTGAA |
R: CTTCAACGCTTCGTGTAACACCAC | R: GCGTCACTCCATCAGGGTTT | ||
Actin | F: CAAAGCGCAAAAAGAACACA | SV-M | F: ACACACTCCACAGTTTACCACCAT |
R: AGAGGAGAGGGCGAGGTTAG | R: CGCCCTCCTGTCAATGAATAG | ||
GFP | F: GTGTTCAATGCTTTGCGAGA | SV-G | F: CCATGTTTCGTTGAGCTTTCC |
R: AAAGGGCAGATTGTGTGGAC | R: CGCCTTCGTGTTCACTGAGTT | ||
TBP | F: TAGTGGCCAATCCTGTGTACCA | SV-L | F: TTCCCTGAAGACGCCCATTA |
R: TCAGCGGAACCTGGTGTGGC | R: TGCCGCCCTCATCCAA |
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Liao, J.-F.; Wu, C.-P.; Tang, C.-K.; Tsai, C.-W.; Rouhová, L.; Wu, Y.-L. Identification of Regulatory Host Genes Involved in Sigma Virus Replication Using RNAi Knockdown in Drosophila. Insects 2019, 10, 339. https://doi.org/10.3390/insects10100339
Liao J-F, Wu C-P, Tang C-K, Tsai C-W, Rouhová L, Wu Y-L. Identification of Regulatory Host Genes Involved in Sigma Virus Replication Using RNAi Knockdown in Drosophila. Insects. 2019; 10(10):339. https://doi.org/10.3390/insects10100339
Chicago/Turabian StyleLiao, Jen-Fu, Carol-P Wu, Cheng-Kang Tang, Chi-Wei Tsai, Lenka Rouhová, and Yueh-Lung Wu. 2019. "Identification of Regulatory Host Genes Involved in Sigma Virus Replication Using RNAi Knockdown in Drosophila" Insects 10, no. 10: 339. https://doi.org/10.3390/insects10100339