Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cells
Arthropod-borne viruses represent an increasing threat to the global health system, requiring the development of novel and sustainable control strategies to reduce the risk of arboviral infections. RNA interference (RNAi) offers a potential approach to directly prevent viral replication within vecto...
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| Language: | English |
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Elsevier
2025-07-01
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| Series: | Virus Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0168170225000619 |
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| author | Alejandra Centurión Bodunrin Omokungbe Sabrina Stiehler Andreas Vilcinskas Kornelia Hardes |
| author_facet | Alejandra Centurión Bodunrin Omokungbe Sabrina Stiehler Andreas Vilcinskas Kornelia Hardes |
| author_sort | Alejandra Centurión |
| collection | DOAJ |
| description | Arthropod-borne viruses represent an increasing threat to the global health system, requiring the development of novel and sustainable control strategies to reduce the risk of arboviral infections. RNA interference (RNAi) offers a potential approach to directly prevent viral replication within vectors due to its specificity in gene silencing. In this study, we evaluated the efficacy of long double-stranded RNAs (dsRNAs) targeting six regions of the Semliki Forest virus (SFV) genome in Aedes albopictus U4.4 cells. The antiviral efficiency of dsRNA alone is low, therefore we evaluated its use after complexing with the K4 Transfection System (K4). A cytotoxicity assay based on ATP quantification showed that both uncomplexed and complexed dsRNA had no cytotoxic effects on U4.4 cells at a concentration up to 2 ng/µL. Complexed dsRNA achieved higher antiviral efficacy, significantly reducing viral replication compared to uncomplexed dsRNA. We found that complexed dsRNA retained its antiviral activity when challenged with SFV up to 72 h post-transfection. Among our synthesized dsRNA constructs, nsP4-dsRNA in complex with K4 led to an 80 % reduction in viral replication at 72 h post-infection at 0.5 ng/µL. Using RT-qPCR, we confirmed a significant 32.2 % reduction of nsP4 mRNA after transfection of complexed nsP4-dsRNA. Dose response assays showed that complexed dsRNAs with a concentration of 0.5 ng/µL are effective for viral reduction. Our results highlight the importance of efficient dsRNA delivery and selection of critical viral targets, such as nsP4, for successful RNAi-mediated viral suppression. This work elucidates the potential of dsRNAs to target Semliki Forest virus replication, highlighting viral gene targeting as a viable strategy for RNAi-based suppression of arboviral replication. |
| format | Article |
| id | doaj-art-3ebac4cc26b645709a6eae2b199cb85f |
| institution | OA Journals |
| issn | 1872-7492 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Virus Research |
| spelling | doaj-art-3ebac4cc26b645709a6eae2b199cb85f2025-08-20T02:34:10ZengElsevierVirus Research1872-74922025-07-0135719958410.1016/j.virusres.2025.199584Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cellsAlejandra Centurión0Bodunrin Omokungbe1Sabrina Stiehler2Andreas Vilcinskas3Kornelia Hardes4Centre for Translational Biodiversity Genomics (LOEWE TBG), Senckenberganlage 25, 60325 Frankfurt am Main, Germany; Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch of Bioresources, Ohlebergsweg 12, 35392 Giessen, GermanyCentre for Translational Biodiversity Genomics (LOEWE TBG), Senckenberganlage 25, 60325 Frankfurt am Main, Germany; Institute for Insect Biotechnology, Justus-Liebig University, Heinrich-Buff-Ring 26-32, 35392 Giessen, GermanyInstitute for Insect Biotechnology, Justus-Liebig University, Heinrich-Buff-Ring 26-32, 35392 Giessen, GermanyCentre for Translational Biodiversity Genomics (LOEWE TBG), Senckenberganlage 25, 60325 Frankfurt am Main, Germany; Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch of Bioresources, Ohlebergsweg 12, 35392 Giessen, Germany; Institute for Insect Biotechnology, Justus-Liebig University, Heinrich-Buff-Ring 26-32, 35392 Giessen, GermanyCentre for Translational Biodiversity Genomics (LOEWE TBG), Senckenberganlage 25, 60325 Frankfurt am Main, Germany; Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch of Bioresources, Ohlebergsweg 12, 35392 Giessen, Germany; BMBF Junior Research Group in Infection Research ''ASCRIBE'', Ohlebergsweg 12, 35392, Giessen, Germany; Corresponding author at: Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Branch of Bioresources, Ohlebergsweg 12, 35392 Giessen, Germany.Arthropod-borne viruses represent an increasing threat to the global health system, requiring the development of novel and sustainable control strategies to reduce the risk of arboviral infections. RNA interference (RNAi) offers a potential approach to directly prevent viral replication within vectors due to its specificity in gene silencing. In this study, we evaluated the efficacy of long double-stranded RNAs (dsRNAs) targeting six regions of the Semliki Forest virus (SFV) genome in Aedes albopictus U4.4 cells. The antiviral efficiency of dsRNA alone is low, therefore we evaluated its use after complexing with the K4 Transfection System (K4). A cytotoxicity assay based on ATP quantification showed that both uncomplexed and complexed dsRNA had no cytotoxic effects on U4.4 cells at a concentration up to 2 ng/µL. Complexed dsRNA achieved higher antiviral efficacy, significantly reducing viral replication compared to uncomplexed dsRNA. We found that complexed dsRNA retained its antiviral activity when challenged with SFV up to 72 h post-transfection. Among our synthesized dsRNA constructs, nsP4-dsRNA in complex with K4 led to an 80 % reduction in viral replication at 72 h post-infection at 0.5 ng/µL. Using RT-qPCR, we confirmed a significant 32.2 % reduction of nsP4 mRNA after transfection of complexed nsP4-dsRNA. Dose response assays showed that complexed dsRNAs with a concentration of 0.5 ng/µL are effective for viral reduction. Our results highlight the importance of efficient dsRNA delivery and selection of critical viral targets, such as nsP4, for successful RNAi-mediated viral suppression. This work elucidates the potential of dsRNAs to target Semliki Forest virus replication, highlighting viral gene targeting as a viable strategy for RNAi-based suppression of arboviral replication.http://www.sciencedirect.com/science/article/pii/S0168170225000619RNA interferenceArbovirusAlphavirusU4.4Cell cultureMosquito |
| spellingShingle | Alejandra Centurión Bodunrin Omokungbe Sabrina Stiehler Andreas Vilcinskas Kornelia Hardes Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cells Virus Research RNA interference Arbovirus Alphavirus U4.4 Cell culture Mosquito |
| title | Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cells |
| title_full | Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cells |
| title_fullStr | Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cells |
| title_full_unstemmed | Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cells |
| title_short | Inhibition of Semliki Forest virus replication with long double-stranded RNA in Aedes albopictus cells |
| title_sort | inhibition of semliki forest virus replication with long double stranded rna in aedes albopictus cells |
| topic | RNA interference Arbovirus Alphavirus U4.4 Cell culture Mosquito |
| url | http://www.sciencedirect.com/science/article/pii/S0168170225000619 |
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