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Nat Biotechnol. 2018 Oct;36(9):880-887. doi: 10.1038/nbt.4201. Epub 2018 Aug 20.

Efficient proximity labeling in living cells and organisms with TurboID.

Author information

1
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
Departments of Genetics, Stanford University, Stanford, California, USA.
3
Department of Chemistry, Stanford University, Stanford, California, USA.
4
Department of Biology, Stanford University, Stanford, California, USA.
5
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
6
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
7
Howard Hughes Medical Institute, Boston, Massachusetts, USA.
8
Chan Zuckerberg Biohub, San Francisco, California, USA.

Abstract

Protein interaction networks and protein compartmentalization underlie all signaling and regulatory processes in cells. Enzyme-catalyzed proximity labeling (PL) has emerged as a new approach to study the spatial and interaction characteristics of proteins in living cells. However, current PL methods require over 18 h of labeling time or utilize chemicals with limited cell permeability or high toxicity. We used yeast display-based directed evolution to engineer two promiscuous mutants of biotin ligase, TurboID and miniTurbo, which catalyze PL with much greater efficiency than BioID or BioID2, and enable 10-min PL in cells with non-toxic and easily deliverable biotin. Furthermore, TurboID extends biotin-based PL to flies and worms.

PMID:
30125270
PMCID:
PMC6126969
[Available on 2019-02-20]
DOI:
10.1038/nbt.4201

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