show Abstracthide AbstractThe goal of this project is to leverage Quantitative Insertion-site Sequencing (QIseq) methods, an Illumina sequencing approach to identify piggyBac transposon insertion sites, to experimentally identify and validate critical processes and pathways in different life cycle stages of the major human malaria parasite Plasmodium falciparum. Previously, we combined QIseq with optimized piggyBac transfection to create >30,000 single random insertions across the Plasmodium falciparum genome. This dataset allowed us to discern genes and pathways that are essential and dispensable for intraerythrocytic growth in routine in vitro culture. In this study we are expanding the approach to other stages of the parasite life cycle, specifically those involved in transmission to the mosquito, and also measuring the response of large pools of mutants to drug and other phenotypic pressures such as heat shock. These experiments will generate valuable functional information about the Plasmodium falciparum genome, large portions of which remain unstudied, and therefore will provide a valuable resource to the broader community.