A non-invasive method for silencing gene transcription in honeybees maintained under natural conditions


Autoria(s): Franco Nunes, Francis Morais; Paulino Simoes, Zila Luz
Contribuinte(s)

Universidade Estadual Paulista (UNESP)

Data(s)

20/05/2014

20/05/2014

01/02/2009

Resumo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Processo FAPESP: 05/03926-5

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 mu g of dsVg-RNA or dsGFP-RNA was offered while no larvae that received 3.0 mu g 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. (c) 2008 Elsevier Ltd. All rights reserved.

Formato

157-160

Identificador

http://dx.doi.org/10.1016/j.ibmb.2008.10.011

Insect Biochemistry and Molecular Biology. Oxford: Pergamon-Elsevier B.V. Ltd, v. 39, n. 2, p. 157-160, 2009.

0965-1748

http://hdl.handle.net/11449/1118

10.1016/j.ibmb.2008.10.011

WOS:000264608200011

Idioma(s)

eng

Publicador

Pergamon-Elsevier B.V. Ltd

Relação

Insect Biochemistry and Molecular Biology

Direitos

closedAccess

Palavras-Chave #Apis mellifera #Honey bee #Vitellogenin #RNAi feeding #Gene knockdown #Development #Non-invasive method #Off-target effects #Environmental RNAi
Tipo

info:eu-repo/semantics/article