Targeted Protein Degradation Using DeGradFP in

Overview

Journal Wellcome Open Res

Specialty Biomedical Engineering

Date 2022 Oct 27

PMID 35865221

Authors

Midori Ishii

Bungo Akiyoshi

Affiliations

Soon will be listed here.

Abstract

Targeted protein degradation is an invaluable tool in studying the function of proteins. Such a tool was not available in , 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 . 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 , deGradFP can serve as a powerful tool to rapidly deplete proteins of interest, especially those with low turnover rates.

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