Research article

piggyBac is an effective tool for functional analysis of the Plasmodium falciparum genome

Bharath Balu¹ , Chitra Chauhan¹ , Steven P Maher¹ , Douglas A Shoue² , Jessica C Kissinger^3,4 , Malcolm J Fraser Jr² and John H Adams¹

¹  Department of Global Health, 3720 Spectrum Blvd Suite 304, University of South Florida, Tampa, Florida 33612, USA

²  Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA

³  Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, Georgia 30602, USA

⁴  Department of Genetics, University of Georgia, Athens, Georgia 30602, USA

author email corresponding author email

BMC Microbiology 2009, 9:83doi:10.1186/1471-2180-9-83

Published:	7 May 2009

Abstract

Background

Much of the Plasmodium falciparum genome encodes hypothetical proteins with limited homology to other organisms. A lack of robust tools for genetic manipulation of the parasite limits functional analysis of these hypothetical proteins and other aspects of the Plasmodium genome. Transposon mutagenesis has been used widely to identify gene functions in many organisms and would be extremely valuable for functional analysis of the Plasmodium genome.

Results

In this study, we investigated the lepidopteran transposon, piggyBac, as a molecular genetic tool for functional characterization of the Plasmodium falciparum genome. Through multiple transfections, we generated 177 unique P. falciparum mutant clones with mostly single piggyBac insertions in their genomes. Analysis of piggyBac insertion sites revealed random insertions into the P. falciparum genome, in regards to gene expression in parasite life cycle stages and functional categories. We further explored the possibility of forward genetic studies in P. falciparum with a phenotypic screen for attenuated growth, which identified several parasite genes and pathways critical for intra-erythrocytic development.

Conclusion

Our results clearly demonstrate that piggyBac is a novel, indispensable tool for forward functional genomics in P. falciparum that will help better understand parasite biology and accelerate drug and vaccine development.