Malaria pp 161-176

Part of the Methods in Molecular Biology book series (MIMB, volume 923)

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

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 

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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