A general strategy to construct small molecule biosensors in eukaryotes
Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Here, we produce biosensors based on a ligand-binding domain (LBD) by using a method that, in principle, can be applied to any target molecule. The LBD is fused to either a fluorescent protein or a transcriptional activator and is destabilized by mutation such that the fusion accumulates only in cells containing the target ligand. We illustrate the power of this method by developing biosensors for digoxin and progesterone. Addition of ligand to yeast, mammalian or plant cells expressing a biosensor activates transcription with a dynamic range of up to ~100-fold. We use the biosensors to improve the biotransformation of pregnenolone to progesterone in yeast and to regulate CRISPR activity in mammalian cells. This work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes.
Article and author information
- Jeffery W Kelly, Scripps Research Institute, United States
- Received: August 6, 2015
- Accepted: December 17, 2015
- Accepted Manuscript published: December 29, 2015 (version 1)
- Accepted Manuscript updated: December 30, 2015 (version 2)
- Version of Record published: January 26, 2016 (version 3)
© 2015, Feng et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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