Liquid-liquid phase separation of light-inducible transcription factors increases transcription activation in mammalian cells and mice
Open Access
- 1 January 2021
- journal article
- research article
- Published by American Association for the Advancement of Science (AAAS) in Science Advances
- Vol. 7 (1), eabd3568
- https://doi.org/10.1126/sciadv.abd3568
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.Keywords
Funding Information
- Deutsche Forschungsgemeinschaft (EXC-294, EXC-2189 Project ID: 390939984, GSC-4, DFG WE 4733/7-1)
- Deutsche Forschungsgemeinschaft (EXC-294 and EXC-2189 - Project ID: 390939984)
- National Natural Science Foundation of China (no. 31971346, no.31861143016)
- Science and Technology Commission of Shanghai Municipality (No. 18JC1411000)
- National Key Laboratory Foundation of China (no. 2019YFA0904500)
This publication has 55 references indexed in Scilit:
- Using optogenetics to tackle systems-level questions of multicellular morphogenesisCurrent Opinion in Cell Biology, 2020
- Principles and applications of optogenetics in developmental biologyDevelopment, 2019
- Regulation of Cellular Metabolism through Phase Separation of EnzymesBiomolecules, 2018
- Light‐Controlled Mammalian Cells and Their Therapeutic Applications in Synthetic BiologyAdvanced Science, 2018
- OptoBase: A Web Platform for Molecular OptogeneticsACS Synthetic Biology, 2018
- Synthetic biological approaches to optogenetically control cell signalingCurrent Opinion in Biotechnology, 2017
- Stress-Triggered Phase Separation Is an Adaptive, Evolutionarily Tuned ResponseCell, 2017
- A bacterial phytochrome-based optogenetic system controllable with near-infrared lightNature Methods, 2016
- A Synthetic Optogenetic Transcription Device Enhances Blood-Glucose Homeostasis in MiceScience, 2011
- Germline P Granules Are Liquid Droplets That Localize by Controlled Dissolution/CondensationScience, 2009