CellFIE: CRISPR- and Cell Fusion-based Two-hybrid Interaction Mapping of Endogenous Proteins

Christopher Secker; Simona Kostova; Hannah Niederlechner; Stephanie Beetz; Ina Wendland; Mara J Liebich; Oliver Polzer; Mirjam Groh; Sigrid Schnoegl; Philipp Trepte; Erich E Wanker

doi:10.1016/j.jmb.2021.167305

CellFIE: CRISPR- and Cell Fusion-based Two-hybrid Interaction Mapping of Endogenous Proteins

J Mol Biol. 2021 Dec 3;433(24):167305. doi: 10.1016/j.jmb.2021.167305. Epub 2021 Oct 14.

Affiliations

¹ Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; Department of Neurology, Charité Universitätsmedizin Berlin, Berlin, Germany. Electronic address: https://twitter.com/Chris_Secker.
² Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
³ Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; Department of Neurology, Charité Universitätsmedizin Berlin, Berlin, Germany.
⁴ Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; Brain Development and Disease, Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Vienna, Austria. Electronic address: philipp.trepte@imba.oeaw.ac.at.
⁵ Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany. Electronic address: erich.w@mdc-berlin.de.

PMID: 34655654
DOI: 10.1016/j.jmb.2021.167305

Abstract

Numerous genetic methods facilitate the detection of binary protein-protein interactions (PPIs) by exogenous overexpression, which can lead to false results. Here, we describe CellFIE, a CRISPR- and cell fusion-based PPI detection method, which enables the mapping of interactions between endogenously tagged two-hybrid proteins. We demonstrate the specificity and reproducibility of CellFIE in a matrix mapping approach, validating the interactions of VCP with ASPL and UBXD1, and the self-interaction of TDP-43 under endogenous conditions. Furthermore, we show that CellFIE can be used to quantify changes of endogenous PPIs upon stress induction or drug treatment. For the first time, CellFIE facilitates systematic mapping of interactions between endogenously tagged proteins and represents a novel tool to characterize PPIs in live cells under dynamic conditions.

Keywords: BRET; cell fusion; endogenous tagging; protein–protein interactions (PPIs); two-hybrid.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Fusion*
Clustered Regularly Interspaced Short Palindromic Repeats*
Humans
Protein Interaction Mapping / methods*
Two-Hybrid System Techniques