TY - JOUR T1 - Targeted protein degradation using degradFP in <em>Trypanosoma brucei</em> JF - bioRxiv DO - 10.1101/2022.06.01.494430 SP - 2022.06.01.494430 AU - Midori Ishii AU - Bungo Akiyoshi Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/06/02/2022.06.01.494430.abstract N2 - Targeted protein degradation is an invaluable tool in studying the function of proteins. Such a tool was not available in Trypanosoma brucei, an evolutionarily divergent eukaryote that causes human African trypanosomiasis. Here, we have adapted degradFP (degrade Green Fluorescent Protein), a protein degradation system based on the SCF E3 ubiquitin ligase complex and anti-GFP nanobody, in T. brucei. As a proof of principle, we targeted a kinetoplastid kinetochore protein (KKT3) that constitutively localizes at kinetochores in the nucleus. Induction of degradFP in a cell line that had both alleles of KKT3 tagged with YFP caused more severe growth defect than RNAi in procyclic (insect form) cells. degradFP also worked on a cytoplasmic protein (COPII subunit, SEC31). Given the easiness in making GFP fusion cell lines in T. brucei, degradFP can serve as a powerful tool to rapidly deplete proteins of interest, especially those with low turnover rates.Competing Interest StatementThe authors have declared no competing interest. ER -