An auxin-inducible degron system for trypanosomes

Abstract

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.