Malaria pp 161-176 | Cite as

Silencing of Genes and Alleles by RNAi in Anopheles gambiae

  • Marina Lamacchia
  • John R. Clayton
  • Rui Wang-Sattler
  • Lars M. Steinmetz
  • Elena A. Levashina
  • Stéphanie A. Blandin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 923)

Abstract

Anopheles gambiae mosquitoes are the major vectors of human malaria parasites. However, mosquitoes are not passive hosts for parasites, actively limiting their development in vivo. Our current understanding of the mosquito antiparasitic response is mostly based on the phenotypic analysis of gene knockdowns obtained by RNA interference (RNAi), through the injection or transfection of long dsRNAs in adult mosquitoes or cultured cells, respectively. Recently, RNAi has been extended to silence specifically one allele of a given gene in a heterozygous context, thus allowing to compare the contribution of different alleles to a phenotype in the same genetic background.

Key words

Malaria Mosquito RNAi Reciprocal allele-specific RNAi Real-time PCR Western blot 

Notes

Acknowledgments

The authors acknowledge the continuous support and interest of Professor Jules A. Hoffmann and members of the laboratories in Strasbourg and Heidelberg for constructive discussions. This work was supported by grants from CNRS, INSERM E.A.L. and S.A.B, by a European Research Council Starting Grant (S.A.B), by the Seventh European Commission Programme “Network of Excellence” Evimalar (E.A.L), and by grants from NIH and the Deutsche Forschunggemeinschaft (L.S.M).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Marina Lamacchia
    • 1
  • John R. Clayton
    • 1
  • Rui Wang-Sattler
    • 2
  • Lars M. Steinmetz
    • 3
  • Elena A. Levashina
    • 1
  • Stéphanie A. Blandin
    • 1
  1. 1.Institut de Biologie Moléculaire et CellulaireStrasbourgFrance
  2. 2.Research Unit of Molecular Epidemiology, Helmholtz Zentrum MünchenNeuherbergGermany
  3. 3.Genome Biology Unit, European Molecular Biology LaboratoryHeidelbergGermany

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