title: A genetic system to study Plasmodium falciparum protein function.
abstract: Current systems to study essential genes in the human malaria parasite Plasmodium falciparum are often inefficient and time intensive, and they depend on the genetic modification of the target locus, a process hindered by the low frequency of integration of episomal DNA into the genome. Here, we introduce a method, termed selection-linked integration (SLI), to rapidly select for genomic integration. SLI allowed us to functionally analyze targets at the gene and protein levels, thus permitting mislocalization of native proteins, a strategy known as knock sideways, floxing to induce diCre-based excision of genes and knocking in altered gene copies. We demonstrated the power and robustness of this approach by validating it for more than 12 targets, including eight essential ones. We also localized and inducibly inactivated Kelch13, the protein associated with artemisinin resistance. We expect this system to be widely applicable for P. falciparum and other organisms with limited genetic tractability.
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Jakob
authors.initials: J
authors.lastname: Birnbaum
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Flemming
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Nick
authors.initials: N
authors.lastname: Reichard
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Alexandra Blancke
authors.initials: AB
authors.lastname: Soares
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Paolo
authors.initials: P
authors.lastname: Mesén-Ramírez
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Ernst
authors.initials: E
authors.lastname: Jonscher
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Bärbel
authors.initials: B
authors.lastname: Bergmann
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Parasitology Section, Hamburg, Germany.
authors.forename: Tobias
authors.identifier: http://orcid.org/0000-0002-3968-4601
authors.initials: T
authors.lastname: Spielmann
mesh_terms: D037621:Artemisinins / Q000494:pharmacology; D004351:Drug Resistance / Q000187:drug effects / Q000235:genetics; D005816:Genetic Complementation Test; D005821:Genetic Techniques; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020696:rab5 GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D037621:Artemisinins; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C031327:artemisinin; D020696:rab5 GTP-Binding Proteins