Targeted Mutagenesis in the Malaria Mosquito Using TALE Nucleases

PLoS One. 2013 Aug 15;8(8):e74511. doi: 10.1371/journal.pone.0074511. eCollection 2013.

Abstract

Anopheles gambiae, the main mosquito vector of human malaria, is a challenging organism to manipulate genetically. As a consequence, reverse genetics studies in this disease vector have been largely limited to RNA interference experiments. Here, we report the targeted disruption of the immunity gene TEP1 using transgenic expression of Transcription-Activator Like Effector Nucleases (TALENs), and the isolation of several TEP1 mutant A. gambiae lines. These mutations inhibited protein production and rendered TEP1 mutants hypersusceptible to Plasmodium berghei. The TALEN technology opens up new avenues for genetic analysis in this disease vector and may offer novel biotechnology-based approaches for malaria control.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Anopheles / genetics*
  • Anopheles / parasitology*
  • Base Sequence
  • Endonucleases / metabolism*
  • Gene Targeting*
  • Genes, Insect
  • Humans
  • Insect Proteins / chemistry
  • Insect Proteins / genetics
  • Malaria / parasitology*
  • Molecular Sequence Data
  • Mutagenesis / genetics*
  • Mutant Proteins / chemistry
  • Mutant Proteins / genetics
  • Mutation / genetics
  • Plasmodium berghei / physiology
  • Trans-Activators / metabolism*

Substances

  • Insect Proteins
  • Mutant Proteins
  • TEP1 protein, Anopheles gambiae
  • Trans-Activators
  • Endonucleases

Grant support

This study was funded by an Agence Nationale de la Recherche (ANR) young researcher grant to E.M. (project GEMM), by the European Commission FP7 projects INFRAVEC (grant agreement no. 228421) and EVIMalar (grant agreement no. 242095), by the Institut National de la Santé et de la Recherche Médicale (INSERM) and by the Centre National de la Recherche Scientifique (CNRS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.