Inducible protein expression in stably transfected cells paves the way toward in-cell NMR studies in defined physiological states and 3D tissue cultures.

22 March 2024, Version 1

Abstract

In-cell NMR spectroscopy is the sole technique for characterizing protein structure, dynamics, and interactions in living human cells at atomic resolution. However, its applications have been restricted to asynchronous single-cell suspensions. We developed a strategy based on target protein inducible overexpression in stably transfected cells, allowing the acquisition of high-resolution in-cell NMR spectra in physiologically defined cellular states and 3D tissue models.

Keywords

in-cell NMR
stable cell lines
inducible protein expression
cell cycle synchronization
spheroids
protein
NMR spectroscopy

Supplementary materials

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Title
Inducible protein expression in stably transfected cells paves the way toward in-cell NMR studies in defined physiological states and 3D tissue cultures.
Description
Supplementary Materials contain Supplementary Figures 1 - 5, referenced in the main text.
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