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CellFIE: CRISPR- and Cell Fusion-based Two-hybrid Interaction Mapping of Endogenous Proteins.
- Source :
-
Journal of molecular biology [J Mol Biol] 2021 Dec 03; Vol. 433 (24), pp. 167305. Date of Electronic Publication: 2021 Oct 14. - Publication Year :
- 2021
-
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.<br />Competing Interests: Conflict of interest The authors declare that they have no conflict of interest.<br /> (Copyright © 2021 Elsevier Ltd. All rights reserved.)
Details
- Language :
- English
- ISSN :
- 1089-8638
- Volume :
- 433
- Issue :
- 24
- Database :
- MEDLINE
- Journal :
- Journal of molecular biology
- Publication Type :
- Academic Journal
- Accession number :
- 34655654
- Full Text :
- https://doi.org/10.1016/j.jmb.2021.167305