RT Journal Article
SR Electronic
T1 Liquid-liquid phase separation of light-inducible transcription factors increases transcription activation in mammalian cells and mice
JF Science Advances
JO Sci Adv
FD American Association for the Advancement of Science
SP eabd3568
DO 10.1126/sciadv.abd3568
VO 7
IS 1
A1 Schneider, Nils
A1 Wieland, Franz-Georg
A1 Kong, Deqiang
A1 Fischer, Alexandra A. M.
A1 Hörner, Maximilian
A1 Timmer, Jens
A1 Ye, Haifeng
A1 Weber, Wilfried
YR 2021
UL http://advances.sciencemag.org/content/7/1/eabd3568.abstract
AB 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.