A Genetic Screen for Attenuated Growth Identifies Genes Crucial
for Intraerythrocytic Development of Plasmodium falciparum
Background
• Malaria is the most devastating human
parasitic infection, threatening half of the
world’s population, & killing more than 1
million people each year
• It’s caused by the parasite Plasmodium, of
which there are 4 species, transmitted by
the female Anopheles mosquito
• Plasmodium falciparum causes the most
severe form of malaria & is responsible for
much of the death associated with the
disease
• Plasmoduim has a very complex life-cycle
with several different stages that make it
difficult for a vaccine, & even drugs, to be
developed
Introduction
• A major obstacle to drug & vaccine
development has been the poor
understanding of the higher value targets
critical in the parasite’s biology
• A considerable amount of the P. falciparum
genome codes for genes that are conserved
among Plasmodium
• Such conserved genes could provide most
information about the parasite’s biology, &
are the best targets for anti-malarial therapy
• Genetic manipulation of P. falciparum is a
challenging process due to a very low
transfection efficiency (the ability of the
parasite to carry non-integrated plasmids as
episomes and the inefficiency to recombine
with the parasite genome)
• Development of a highly efficient,
piggyBac transposon-based mutagenesis
system for P. falciparum, a large-scale,
genome-wide insertion mutagenesis
approach could aid greatly in dissecting the
P. falciparum genome
• Malaria parasite growth in the
intraerythrocytic stages is determined by
multiple factors, each of which represents
the net result of numerous molecular
functions, & may be possible targets for
therapeutic intervention
• Performing an in vitro proliferation assay
for 123 piggyBac- transformed P. falciparum
cloned lines can be used to identify genes
and pathways vital for intraerythrocytic
development
Methods/Results Conclusion
• Forward functional genomics
approaches such as transposon-mediated
insertion mutagenesis could provide
valuable information about the parasite
genome by associating a gene with its
function, while perhaps more
importantly deciphering essential from
non-essential components of the genome
• Apicoplast and mitochondrial genes
provide excellent targets for anti-malarial
therapy, as they are unique to the
parasite
• Four apicoplast genes cause
considerable attenuation of parasite
growth and provide grounds for further
evaluation as anti-malarial targets
• A forward genetics approach can be
used to identify several genes that are
extremely crucial for intraerythrocytic
development of P. falciparum, & provide
an experimental basis for their further
investigation as putative anti-malarial
targets
• Other phenotypic screens for
morphology, virulence, drug resistance,
gametocytogenesis & transmission to
mosquito hosts could be addressed with
the mutant library
References
Balu B., Singh N., Maher S.P., Adams J.H.
et. al. “A Genetic Screen for Attenuated
Growth Identifies Genes Crucial for
Intraerythrocytic Development of
Plasmodium falciparum.” PLoS ONE
5(10): e13282.
doi:10.1371/journal.pone.0013282. 11
Oct 2010.