학술논문
CellFIE: CRISPR- and Cell Fusion-based Two-hybrid Interaction Mapping of Endogenous Proteins.
Document Type
Academic Journal
Author
Secker C; 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.; Kostova S; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.; Niederlechner H; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; Department of Neurology, Charité Universitätsmedizin Berlin, Berlin, Germany.; Beetz S; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.; Wendland I; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.; Liebich MJ; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.; Polzer O; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.; Groh M; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany; Department of Neurology, Charité Universitätsmedizin Berlin, Berlin, Germany.; Schnoegl S; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.; Trepte P; 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.; Wanker EE; Neuroproteomics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany. Electronic address: erich.w@mdc-berlin.de.
Source
Publisher: Elsevier Country of Publication: Netherlands NLM ID: 2985088R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1089-8638 (Electronic) Linking ISSN: 00222836 NLM ISO Abbreviation: J Mol Biol Subsets: MEDLINE
Subject
Language
English
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.
Competing Interests: Conflict of interest The authors declare that they have no conflict of interest.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)
Competing Interests: Conflict of interest The authors declare that they have no conflict of interest.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)