A general strategy to construct small molecule biosensors in eukaryotes

Harvard Medical School, United States
University of Washington, United States
Colorado State University, United States
Howard Hughes Medical Institute, University of Washington, United States

Research Article Dec 29, 2015

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Cite this article as: eLife 2015;4:e10606 doi: 10.7554/eLife.10606

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Abstract

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

Author details

Justin Feng

Program in Biological and Biomedical Sciences, Harvard Medical School, Boston, United States

Competing interests
Justin Feng, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work.
Benjamin W Jester

Department of Genome Sciences, University of Washington, Seattle, United States

Competing interests
Benjamin W Jester, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work.
Christine E Tinberg

Department of Biochemistry, University of Washington, Seattle, United States

Competing interests
Christine E Tinberg, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work.
Daniel J Mandell

Department of Genetics, Harvard Medical School, Boston, United States

For correspondence
djmandell@gmail.com

Competing interests
Daniel J Mandell, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work. Harvard University has filed a provisional patent.
Mauricio S Antunes

Department of Biology, Colorado State University, Fort Collins, United States

Competing interests
No competing interests declared.
Raj Chari

Department of Genetics, Harvard Medical School, Boston, United States

Competing interests
Raj Chari, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work.
Kevin J Morey

Department of Biology, Colorado State University, Fort Collins, United States

Competing interests
No competing interests declared.
Xavier Rios

Department of Genetics, Harvard Medical School, Boston, United States

Competing interests
No competing interests declared.
June I Medford

Department of Biology, Colorado State University, Fort Collins, United States

Competing interests
No competing interests declared.
George M Church

Department of Genetics, Harvard Medical School, Boston, United States

Competing interests
George M Church, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work.
Stanley Fields

Department of Genome Sciences, University of Washington, Seattle, United States

Competing interests
Stanley Fields, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work.
David Baker

Howard Hughes Medical Institute, University of Washington, Seattle, United States

Competing interests
David Baker, has filed a provisional application (patent application number 62220628) with the US Patent and Trademark Office on this work.

Reviewing Editor

Jeffery W Kelly, Scripps Research Institute, United States

Publication history

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)

Copyright

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.