Helmholtz Gemeinschaft

Search
Browse
Statistics
Feeds

CellFIE: CRISPR- and cell fusion-based two-hybrid interaction mapping of endogenous proteins

[thumbnail of Accepted Manuscript (final draft) incl. Supplementary Material]
Preview
PDF (Accepted Manuscript (final draft) incl. Supplementary Material) - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
9MB

Item Type:Article
Title:CellFIE: CRISPR- and cell fusion-based two-hybrid interaction mapping of endogenous proteins
Creators Name:Secker, C., Kostova, S., Niederlechner, H., Beetz, S., Wendland, I., Liebich, M.J., Polzer, O., Groh, M., Schnoegl, S., Trepte, P. and Wanker, E.E.
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:Cell Fusion, Clustered Regularly Interspaced Short Palindromic Repeats, Protein Interaction Mapping, Two-Hybrid System Techniques
Source:Journal of Molecular Biology
ISSN:0022-2836
Publisher:Elsevier
Volume:433
Number:24
Page Range:167305
Date:3 December 2021
Additional Information:Copyright © 2021. This manuscript version is made available under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/ or send a letter to Creative Commons, PO Box 1866, Mountain View, CA 94042, USA.
Official Publication:https://doi.org/10.1016/j.jmb.2021.167305
PubMed:View item in PubMed

Repository Staff Only: item control page

Downloads

Downloads per month over past year

Open Access
MDC Library