An Auxin-Inducible Degron System for Trypanosomes

The ability to specifically degrade one protein in a cell provides an immediate insight into any resulting phenotype and thus function that may be occluded by secondary effects using other techniques. The auxin-inducible degron is a method developed to achieve this end and is effective in mammalian and yeast cells. The approach is dependent on the recognition of a tagged target protein by an endogenous E3 ubiquitin ligase modified by the expression of a plant F-box protein. This approach was initially unsuccessful in trypanosomes, probably due to a lack of interaction between the endogenous E3 ubiquitin ligase and the plant F-box protein. This was overcome by expressing the entire rice E3 ubiquitin ligase in trypanosomes, and here we describe the production of the auxin-inducible degron competent cell lines and tagging of target genes.