Liquid-liquid Phase Separation of Light-inducible Transcription Factors Increases Transcription Activation in Mammalian Cells and Mice

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2021 Feb 1

PMID 33523844

Citations 49

Authors

Nils Schneider

Franz-Georg Wieland

Deqiang Kong

Alexandra A M Fischer

Maximilian Horner

Jens Timmer

Haifeng Ye

Wilfried Weber

Affiliations

Soon will be listed here.

Abstract

Light-inducible gene switches represent a key strategy for the precise manipulation of cellular events in fundamental and applied research. However, the performance of widely used gene switches is limited due to low tissue penetrance and possible phototoxicity of the light stimulus. To overcome these limitations, we engineer optogenetic synthetic transcription factors to undergo liquid-liquid phase separation in close spatial proximity to promoters. Phase separation of constitutive and optogenetic synthetic transcription factors was achieved by incorporation of intrinsically disordered regions. Supported by a quantitative mathematical model, we demonstrate that engineered transcription factor droplets form at target promoters and increase gene expression up to fivefold. This increase in performance was observed in multiple mammalian cells lines as well as in mice following in situ transfection. The results of this work suggest that the introduction of intrinsically disordered domains is a simple yet effective means to boost synthetic transcription factor activity.

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