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| Item Type: | Article |
|---|---|
| Title: | CellFIE: CRISPR- and cell fusion-based two-hybrid interaction mapping of endogenous proteins |
| Creators Name: | Secker, C. and Kostova, S. and Niederlechner, H. and Beetz, S. and Wendland, I. and Liebich, M.J. and Polzer, O. and Groh, M. and Schnoegl, S. and 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 |
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