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To develop effective malaria transmission-blocking vaccines and drugs, it is crucial to understand the genetic factors and molecular mechanisms that regulate the development of Plasmodium blood-stage sexual forms, known as gametocytes—parasite stage capable of surviving in the mosquito vector. We established a scalable forward genetic screen approach using single-insertion mutants generated by random piggyBac mutagenesis. This method identifies genes essential for asexual parasite forms survival or tolerance to critical in vivo phenotype responses, such as febrile temperature, antimalarial drugs, and oxidative stress. Building on this well-established approach, we developed a screen for gametocyte-related phenotypes, categorizing genes based on their impact on gametocyte production and development as either hypo-producers (reduced gametocyte production) or hyper-producers (increased gametocyte production). This approach identifies the genetic factors driving gametocyte conversion and growth. Here, we present the methodology of our large-scale phenotypic screen for identifying essential Plasmodium falciparum gametocyte genes.

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Large-Scale Mutagenesis Screening for Genetic Determinants of Plasmodium falciparum Sexual Development

  • 实验方案

  • 发表日期：

    2026年

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  • Olatunbosun Aringbangba,
  • Camilla Valente Pires,
  • Jyotsna Chawla,
  • Prem Prakash,
  • Lauriane Sollelis,
  • Matthias Marti,
  • John H. Adams

摘要

To develop effective malaria transmission-blocking vaccines and drugs, it is crucial to understand the genetic factors and molecular mechanisms that regulate the development of Plasmodium blood-stage sexual forms, known as gametocytes—parasite stage capable of surviving in the mosquito vector. We established a scalable forward genetic screen approach using single-insertion mutants generated by random piggyBac mutagenesis. This method identifies genes essential for asexual parasite forms survival or tolerance to critical in vivo phenotype responses, such as febrile temperature, antimalarial drugs, and oxidative stress. Building on this well-established approach, we developed a screen for gametocyte-related phenotypes, categorizing genes based on their impact on gametocyte production and development as either hypo-producers (reduced gametocyte production) or hyper-producers (increased gametocyte production). This approach identifies the genetic factors driving gametocyte conversion and growth. Here, we present the methodology of our large-scale phenotypic screen for identifying essential Plasmodium falciparum gametocyte genes.