A non-invasive method for silencing gene transcription in honeybees maintained under natural conditions |
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| Authors: | Francis Morais Franco Nunes Zilá Luz Paulino Simões |
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| Affiliation: | 1. Departamento de Biologia Aplicada a Agropecuária, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal, SP, Brazil;2. Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil;3. Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil;1. USDA Bee Research, 10300 Baltimore Avenue, Building 306, Beltsville, MD 20705, USA;2. Department of Ecology and Evolution, University of Lausanne, CH-1015, Switzerland;3. Department of Entomology, University of Arizona, Tucson, AZ 85721, USA;1. Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil;2. Instituto Federal de São Paulo, Campus Matão, Matão, SP, Brazil;1. Departamento de Biologia Celular e Molecular, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes 3900, 14049-900, Ribeirão Preto, São Paulo, Brazil;2. Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Av. do Café, s/n, 14040-903, Ribeirão Preto, São Paulo, Brazil;1. School of Biological Sciences (Zoology), University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ, UK;2. National Bee Unit, Fera, Sand Hutton, York YO41 1LZ, UK;3. Vita (Europe) Limited, Vita House, London Street, Basingstoke, Hampshire RG21 7PG, UK |
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| Abstract: | In the Apis mellifera post-genomic era, RNAi protocols have been used in functional approaches. However, sample manipulation and invasive methods such as injection of double-stranded RNA (dsRNA) can compromise physiology and survival. To circumvent these problems, we developed a non-invasive method for honeybee gene knockdown, using a well-established vitellogenin RNAi system as a model. Second instar larvae received dsRNA for vitellogenin (dsVg-RNA) in their natural diet. For exogenous control, larvae received dsRNA for GFP (dsGFP-RNA). Untreated larvae formed another control group. Around 60% of the treated larvae naturally developed until adult emergence when 0.5 μg of dsVg-RNA or dsGFP-RNA was offered while no larvae that received 3.0 μg of dsRNA reached pupal stages. Diet dilution did not affect the removal rates. Viability depends not only on the delivered doses but also on the internal conditions of colonies. The weight of treated and untreated groups showed no statistical differences. This showed that RNAi ingestion did not elicit drastic collateral effects. Approximately 90% of vitellogenin transcripts from 7-day-old workers were silenced compared to controls. A large number of samples are handled in a relatively short time and smaller quantities of RNAi molecules are used compared to invasive methods. These advantages culminate in a versatile and a cost-effective approach. |
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