2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum.
Analysis of gene function in Plasmodium falciparum, the most important human malaria parasite, is restricted by the lack of robust and simple reverse genetic tools. Approaches to manipulate protein levels post-translationally are powerful tools to study protein-off effects especially in the haploid malaria parasite where genetic knockouts of essential genes are lethal. We investigated if the auxin-inducible degron system is functional in P. falciparum and found that degron-tagged yellow fluorescent protein levels were efficiently reduced upon addition of auxin which otherwise had no effect on parasite viability. The genetic components required in this conditional approach were co-expressed in P. falciparum by applying the small peptide 2A. 2A is a self-processing peptide from Foot-And-Mouth Disease virus that allows the whole conditional system to be accommodated on a single plasmid vector and ensures stoichiometric expression levels.
Institute of Tropical Medicine, University of Tübingen, Tübingen, Germany.
Andrea
A
Kreidenweiss
Annika V
AV
Hopkins
Benjamin
B
Mordmüller
D001426:Bacterial Proteins / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007210:Indoleacetic Acids / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D008164:Luminescent Proteins / Q000378:metabolism; D010963:Plasmodium falciparum / Q000187:drug effects / Q000378:metabolism*; D059748:Proteolysis / Q000187:drug effects*; D015800:Protozoan Proteins / Q000378:metabolism*; D015533:Transcriptional Activation; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
D001426:Bacterial Proteins; D007210:Indoleacetic Acids; D008164:Luminescent Proteins; D015800:Protozoan Proteins; D014764:Viral Proteins; C054562:yellow fluorescent protein, Bacteria; D003546:Cysteine Endopeptidases