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

Autor/innen

  • C. Secker
  • S. Kostova
  • H. Niederlechner
  • S. Beetz
  • I. Wendland
  • M.J. Liebich
  • O. Polzer
  • M. Groh
  • S. Schnoegl
  • P. Trepte
  • E.E. Wanker

Journal

  • Journal of Molecular Biology

Quellenangabe

  • J Mol Biol 433 (24): 167305

Zusammenfassung

  • 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.


DOI

doi:10.1016/j.jmb.2021.167305