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A method for parental RNA interference in the wasp Nasonia vitripennis

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Abstract

The wasp Nasonia vitripennis is emerging as a useful model organism in which to address a variety of biological questions, due, in part, to its ease of laboratory use, unique aspects of its biology and the sequencing of its genome. In order to take full advantage of the potential of this organism, methods for manipulating gene function are needed. To this end, a protocol for parental RNA interference (pRNAi) in N. vitripennis is described. pRNAi entails injecting pupae with double-stranded RNA, allowing the injected wasps to eclose and examining the progeny for developmental defects. This basic protocol is described in the context of the life cycle of N. vitripennis. This technique has been useful in elucidating the function of most, although not all, genes tested to date, and has potential applications beyond embryonic patterning. pRNAi experiments in Nasonia can be completed in as little as 2 weeks.

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Figure 1: Examples of pupal stages of N. vitripennis
Figure 2: Distinguishing males and females in N. vitripennis
Figure 3: Schematic representation of the general strategy used to produce the template for dsRNA.
Figure 4: An example of wasp pupae affixed to a coverslip, ready for injection.
Figure 5
Figure 6: An example of the components of an egg-laying chamber for collecting N. vitripennis embryos.
Figure 7: Healthy host pupa (S. bullata)
Figure 8: Collecting embryos from a parasitized host.
Figure 9: Dissecting embryos from chorion and vitelline membrane.
Figure 10: Example of effects of pRNAi on Nasonia larval cuticles.
Figure 11: Example of effects of pRNAi on embryonic gene expression.

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Acknowledgements

We would like to thank Mary Anne Pultz and David Leaf for providing invaluable knowledge and assistance in working with Nasonia. This work was supported by a NIH grant to C.D.; J.A.L. was supported by a NIH Training Grant and a Dean Dissertation Fellowship from NYU. This investigation was conducted in a facility constructed with support from a Research Facilities Improvement grant from NCRR, NIH.

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  1. Department of Biology, New York University, 100 Washington Square East, New York, 10003, New York, USA

    Jeremy A Lynch & Claude Desplan

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  1. Jeremy A Lynch
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  2. Claude Desplan
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Correspondence to Claude Desplan.

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Lynch, J., Desplan, C. A method for parental RNA interference in the wasp Nasonia vitripennis. Nat Protoc 1, 486–494 (2006). https://doi.org/10.1038/nprot.2006.70

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