A practical approach to RNA interference for studying gene function in a refractory social insect (on a limited budget).

Autor: Walton, A., Flores, E., Guinness, A., Fortune, R., Sheehan, M. J., Toth, A. L.
Zdroj: Insectes Sociaux; Jun2023, Vol. 70 Issue 2, p213-224, 12p
Abstrakt: With powerful functional genomics tools, including RNA interference (RNAi), the study of gene function is moving from simpler phenotypes in model organisms to more complex traits in non-models. However, optimizing RNAi-mediated gene knockdown for a novel system can be time consuming and expensive, so researchers interested in initiating an RNAi study for functional genomics in a non-model organism will want to consider the successes and failures of RNAi in related species and similar phenotypes. Yet, sub-optimal and negative knockdown results are rarely reported alongside positive ones, so choosing the best methodological parameters can be difficult. Furthermore, social insects pose extra challenges to designing functional genomics studies, as the traits we measure are often part of the extended phenotype of a primary reproductive, emerge from the collective phenotypes of many individuals, and may be the result of a gene taking on a novel functional role in a social system. We attempt to mitigate these challenges for other social insect biologists by reporting our own successes and impediments to optimizing RNAi for the gene vitellogenin in the paper wasp Polistes fuscatus. We discuss the many factors that can affect RNAi experimental design and which parameters worked optimally for our study, as well as compile a list of working parameters from previous RNAi studies in social Hymenoptera. We highlight the difficulties of RNAi that are specific to social Hymenoptera, including the potential differences in RNA efficacy across behavioral caste, and share results on our optimized parameters as a potential roadmap for other social insect biologists. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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