Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi
Abstract Background Insecticides are still at the core of insect pest and vector control programmes. Several lines of evidence indicate that ABC transporters are involved in detoxification processes against insecticides, including permethrin and other pyrethroids. In particular, the ABCG4 gene, a me...
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doaj-2505c408d4da4375b87c97b56538a7b62020-11-25T03:01:11ZengBMCMalaria Journal1475-28752019-08-0118111210.1186/s12936-019-2925-5Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensiAgata Negri0Marco Ferrari1Riccardo Nodari2Edoardo Coppa3Valentina Mastrantonio4Sergio Zanzani5Daniele Porretta6Claudio Bandi7Sandra Urbanelli8Sara Epis9Department of Environmental Biology, Sapienza University of RomeDepartment of Biosciences and Pediatric Clinical Research Center “Romeo ed Enrica Invernizzi”, University of MilanDepartment of Biosciences and Pediatric Clinical Research Center “Romeo ed Enrica Invernizzi”, University of MilanDepartment of Biosciences and Pediatric Clinical Research Center “Romeo ed Enrica Invernizzi”, University of MilanDepartment of Environmental Biology, Sapienza University of RomeDepartment of Veterinary Medicine-DIMEVET, Università degli Studi di MilanoDepartment of Environmental Biology, Sapienza University of RomeDepartment of Biosciences and Pediatric Clinical Research Center “Romeo ed Enrica Invernizzi”, University of MilanDepartment of Environmental Biology, Sapienza University of RomeDepartment of Biosciences and Pediatric Clinical Research Center “Romeo ed Enrica Invernizzi”, University of MilanAbstract Background Insecticides are still at the core of insect pest and vector control programmes. Several lines of evidence indicate that ABC transporters are involved in detoxification processes against insecticides, including permethrin and other pyrethroids. In particular, the ABCG4 gene, a member of the G subfamily, has consistently been shown to be up-regulated in response to insecticide treatments in the mosquito malaria vector Anopheles stephensi (both adults and larvae). Methods To verify the actual involvement of this transmembrane protein in the detoxification process of permethrin, bioassays on larvae of An. stephensi, combining the insecticide with a siRNA, specifically designed for the inhibition of ABCG4 gene expression were performed. Administration to larvae of the same siRNA, labeled with a fluorescent molecule, was effected to investigate the systemic distribution of the inhibitory RNA into the larval bodies. Based on siRNA results, similar experiments using antisense Vivo-Morpholinos (Vivo-MOs) were effected. These molecules, compared to siRNA, are expected to guarantee a higher stability in environmental conditions and in the insect gut, and present thus a higher potential for future in-field applications. Results Bioassays using two different concentrations of siRNA, associated with permethrin, led to an increase of larval mortality, compared with results with permethrin alone. These outcomes confirm that ABCG4 transporter plays a role in the detoxification process against the selected insecticide. Moreover, after fluorescent labelling, it was shown the systemic dissemination of siRNA in different body districts of An. stephensi larvae, which suggest a potential systemic effect of the molecule. At the same time, results of Vivo-MO experiments were congruent with those obtained using siRNA, thus confirming the potential of ABCG4 inhibition as a strategy to increase permethrin susceptibility in mosquitoes. For the first time, Vivo-MOs were administered in water to larvae, with evidence for a biological effect. Conclusions Targeting ABCG4 gene for silencing through both techniques resulted in an increased pyrethroid efficacy. These results open the way toward the possibility to exploit ABCG4 inhibition in the context of integrated programmes for the control An. stephensi mosquitoes and malaria transmission.http://link.springer.com/article/10.1186/s12936-019-2925-5Insecticide detoxificationABC-transporter inhibitionMosquito controlsiRNAVivo-MO |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Agata Negri Marco Ferrari Riccardo Nodari Edoardo Coppa Valentina Mastrantonio Sergio Zanzani Daniele Porretta Claudio Bandi Sandra Urbanelli Sara Epis |
spellingShingle |
Agata Negri Marco Ferrari Riccardo Nodari Edoardo Coppa Valentina Mastrantonio Sergio Zanzani Daniele Porretta Claudio Bandi Sandra Urbanelli Sara Epis Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi Malaria Journal Insecticide detoxification ABC-transporter inhibition Mosquito control siRNA Vivo-MO |
author_facet |
Agata Negri Marco Ferrari Riccardo Nodari Edoardo Coppa Valentina Mastrantonio Sergio Zanzani Daniele Porretta Claudio Bandi Sandra Urbanelli Sara Epis |
author_sort |
Agata Negri |
title |
Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi |
title_short |
Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi |
title_full |
Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi |
title_fullStr |
Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi |
title_full_unstemmed |
Gene silencing through RNAi and antisense Vivo-Morpholino increases the efficacy of pyrethroids on larvae of Anopheles stephensi |
title_sort |
gene silencing through rnai and antisense vivo-morpholino increases the efficacy of pyrethroids on larvae of anopheles stephensi |
publisher |
BMC |
series |
Malaria Journal |
issn |
1475-2875 |
publishDate |
2019-08-01 |
description |
Abstract Background Insecticides are still at the core of insect pest and vector control programmes. Several lines of evidence indicate that ABC transporters are involved in detoxification processes against insecticides, including permethrin and other pyrethroids. In particular, the ABCG4 gene, a member of the G subfamily, has consistently been shown to be up-regulated in response to insecticide treatments in the mosquito malaria vector Anopheles stephensi (both adults and larvae). Methods To verify the actual involvement of this transmembrane protein in the detoxification process of permethrin, bioassays on larvae of An. stephensi, combining the insecticide with a siRNA, specifically designed for the inhibition of ABCG4 gene expression were performed. Administration to larvae of the same siRNA, labeled with a fluorescent molecule, was effected to investigate the systemic distribution of the inhibitory RNA into the larval bodies. Based on siRNA results, similar experiments using antisense Vivo-Morpholinos (Vivo-MOs) were effected. These molecules, compared to siRNA, are expected to guarantee a higher stability in environmental conditions and in the insect gut, and present thus a higher potential for future in-field applications. Results Bioassays using two different concentrations of siRNA, associated with permethrin, led to an increase of larval mortality, compared with results with permethrin alone. These outcomes confirm that ABCG4 transporter plays a role in the detoxification process against the selected insecticide. Moreover, after fluorescent labelling, it was shown the systemic dissemination of siRNA in different body districts of An. stephensi larvae, which suggest a potential systemic effect of the molecule. At the same time, results of Vivo-MO experiments were congruent with those obtained using siRNA, thus confirming the potential of ABCG4 inhibition as a strategy to increase permethrin susceptibility in mosquitoes. For the first time, Vivo-MOs were administered in water to larvae, with evidence for a biological effect. Conclusions Targeting ABCG4 gene for silencing through both techniques resulted in an increased pyrethroid efficacy. These results open the way toward the possibility to exploit ABCG4 inhibition in the context of integrated programmes for the control An. stephensi mosquitoes and malaria transmission. |
topic |
Insecticide detoxification ABC-transporter inhibition Mosquito control siRNA Vivo-MO |
url |
http://link.springer.com/article/10.1186/s12936-019-2925-5 |
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