Protocol

Malaria

Volume 923 of the series Methods in Molecular Biology pp 161-176

Date:

Silencing of Genes and Alleles by RNAi in Anopheles gambiae

  • Marina LamacchiaAffiliated withInstitut de Biologie Moléculaire et Cellulaire
  • , John R. ClaytonAffiliated withInstitut de Biologie Moléculaire et Cellulaire
  • , Rui Wang-SattlerAffiliated withResearch Unit of Molecular Epidemiology, Helmholtz Zentrum München
  • , Lars M. SteinmetzAffiliated withGenome Biology Unit, European Molecular Biology Laboratory
  • , Elena A. LevashinaAffiliated withInstitut de Biologie Moléculaire et Cellulaire
  • , Stéphanie A. BlandinAffiliated withInstitut de Biologie Moléculaire et Cellulaire Email author 

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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