Targeted protein degradation using deGradFP in Trypanosoma brucei

Bungo Akiyoshi*, Midori Ishii

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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 [GFP]), 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 yellow fluorescent protein (YFP) caused a more severe growth defect than RNAi in procyclic (insect form) cells. deGradFP also worked on a cytoplasmic protein (COPII subunit, SEC31). Given the ease 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.

Original languageEnglish
Article number175
Number of pages20
JournalWellcome Open Research
Volume7
Early online date23 Jun 2022
DOIs
Publication statusPublished - 19 Oct 2022

Keywords / Materials (for Non-textual outputs)

  • DeGradFP
  • degron
  • kinetochore
  • kinetoplastid
  • targeted protein degradation
  • trypanosoma brucei

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