A general strategy to construct small molecule biosensors in eukaryotes

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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.

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Justin Feng

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

Competing interests
Harvard University has filed a provisional patent on this work
Benjamin W Jester

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

Competing interests
Harvard University has filed a provisional patent on this work
Christine E Tinberg

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

Competing interests
Harvard University has filed a provisional patent on this work
Daniel J Mandell

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

For correspondence
djmandell@gmail.com

Competing interests
Harvard University has filed a provisional patent on this work
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
Harvard University has filed a provisional patent 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
Harvard University has filed a provisional patent on this work
Stanley Fields

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

Competing interests
Harvard University has filed a provisional patent on this work
David Baker

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

Competing interests
Harvard University has filed a provisional patent on this work

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