Efficient proximity labeling in living cells and organisms with TurboID

被引：1022

作者：

Branon, Tess C. ^{[1
,2
,3
,4
]}

Bosch, Justin A. ^{[5
]}

Sanchez, Ariana D. ^{[4
]}

Udeshi, Namrata D. ^{[6
]}

Svinkina, Tanya ^{[6
]}

Carr, Steven A. ^{[6
]}

Feldman, Jessica L. ^{[4
]}

Perrimon, Norbert ^{[5
,7
]}

Ting, Alice Y. ^{[1
,2
,3
,4
,8
]}

机构：

[1] MIT, Dept Chem, Cambridge, MA 02139 USA

[2] Stanford Univ, Dept Genet, Stanford, CA 94305 USA

[3] Stanford Univ, Dept Chem, Stanford, CA 94305 USA

[4] Stanford Univ, Dept Biol, Stanford, CA 94305 USA

[5] Harvard Med Sch, Dept Genet, Boston, MA USA

[6] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[7] Howard Hughes Med Inst, Boston, MA 02115 USA

[8] Chan Zuckerberg Biohub, San Francisco, CA 94158 USA

来源：

NATURE BIOTECHNOLOGY | 2018年 / 36卷 / 09期

关键词：

CAENORHABDITIS-ELEGANS; BIOTIN REPRESSOR; PROTEIN; PEROXIDASE; IDENTIFICATION; COMPONENTS; REPORTER; TOPOLOGY; KINASE; LIGASE;

D O I：

10.1038/nbt.4201

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：880 / +

页数：19