Tet Trim-Away: A conditional rapid protein degradation system for Tetrahymena thermophila.

Authors

Gayatri L Dholakia, Alexander J Stemm-Wolf, Aaisha Anzar, Aaron P Turkewitz, Chad G Pearson

Published in

Molecular biology of the cell. Pages mbcE26050233. Aug 05, 2026. Epub Aug 05, 2026.

Abstract

Tetrahymena thermophila is a ciliated protist that has played pivotal roles in biological discovery. Functional studies of Tetrahymena proteins have largely relied on gene knockouts. Because protein depletion upon knockout typically spans multiple cell cycles, compensatory mechanisms can confound phenotypic interpretation. To enable rapid and acute protein depletion, we modified and adapted the Trim-Away system for use in Tetrahymena (Tet Trim-Away). Trim-Away is based on the E3 ubiquitin ligase, TRIM21, that binds to antibody-bound proteins and targets them for proteasome mediated degradation. Here, Trim-Away was modified with a fusion of the N-terminal RBCC (RING, B-box, coiled-coil) domains of TRIM21 with an α-mCherry (mCh) nanobody sequence that recognizes endogenously tagged mCh proteins of interest (Nb^mCh). Expression of the RBCC:Nb^mCh degron, which is controlled by an inducible promotor, promotes rapid target protein depletion within 30 minutes and can be sustained for weeks. Tet Trim-Away is reversible, functions against targets in multiple cellular compartments, and produces loss-of-function phenotypes in Tetrahymena cells.

PMID:
42555825
Bibliographic data and abstract were imported from PubMed on 06 Aug 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.