Long-term effects and parental RNAi in the blood feeder Rhodnius prolixus (Hemiptera; Reduviidae) |
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| Authors: | Rafaela MM Paim Ricardo N Araujo Michael J Lehane Nelder F Gontijo Marcos H Pereira |
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| Institution: | 1. Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Bloco I4, Sala 177, Av. Antonio Carlos 6627, Pampulha, CEP 30270-901 Belo Horizonte, MG, Brazil;2. Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK;3. Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, CEP 21941-909 Rio de Janeiro, RJ, Brazil;1. Graduate School of Science, Osaka City University, Osaka, Japan;2. Graduate School of Science, Kyoto University, Kyoto, Japan;1. Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan;2. National Agricultural and Food Research Organization, Ibaraki 305-8634, Japan;3. Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal;4. Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-0032, Japan;5. Department of Biology, University of Copenhagen, Copenhagen 2200, Denmark;6. School of Biological Sciences, Monash University, Victoria 3800, Australia;7. Faculty of Food and Agricultural Sciences, Fukushima University, Fukushima 960-1296, Japan;1. Centro de Bioinvestigaciones and Departamento de Ciencias Básicas y Experimentales, Universidad Nacional del Noroeste de la Provincia de Buenos Aires, Monteagudo 2772, 2700, Pergamino, Buenos Aires, Argentina;2. Laboratorio de Genética y Genómica Funcional, Centro Regional de Estudios Genómicos, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Bvd 120 y 62, La Plata, Buenos Aires, Argentina;3. Centro de Investigaciones sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pública, Av. Universidad 655, Sta. María Ahuacatitlán, Cuernavaca, Mexico |
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| Abstract: | RNA interference (RNAi) has been widely employed as a useful alternative to study gene function in insects, including triatomine bugs. However, several aspects related to the RNAi mechanism and functioning are still unclear. The aim of this study is to investigate the persistence and the occurrence of systemic and parental RNAi in the triatomine bug Rhodnius prolixus. For such, the nitrophorins 1 to 4 (NP1-4), which are salivary hemeproteins, and the rhodniin, an intestinal protein, were used as targets for RNAi. The dsRNA for both molecules were injected separately into 3rd and 5th instar nymphs of R. prolixus and the knockdown (mRNA levels and phenotype) were progressively evaluated along several stages of the insect's life. We observed that the NP1-4 knockdown persisted for more than 7 months after the dsRNA injection, and at least 5 months in rhodniin knockdown, passing through various nymphal stages until the adult stage, without continuous input of dsRNA. The parental RNAi was successful from the dsRNA injection in 5th instar nymphs for both knockdown targets, when the RNAi effects (mRNA levels and phenotype) were observed at least in the 2nd instar nymphs of the F1 generation. However, the parental RNAi did not occur when the dsRNA was injected in the 3rd instars. The confirmation of the long persistence and parental transmission of RNAi in R. prolixus can improve and facilitate the utilization of this tool in insect functional genomic studies. |
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| Keywords: | RNA interference Persistence Parental RNAi |
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