A general strategy to construct small molecule biosensors in eukaryotes

  1. Justin Feng
  2. Benjamin W Jester
  3. Christine E Tinberg
  4. Daniel J Mandell  Is a corresponding author
  5. Mauricio S Antunes
  6. Raj Chari
  7. Kevin J Morey
  8. Xavier Rios
  9. June I Medford
  10. George M Church
  11. Stanley Fields
  12. David Baker
  1. Harvard Medical School, United States
  2. University of Washington, United States
  3. Colorado State University, United States
  4. Howard Hughes Medical Institute, University of Washington, United States

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

  1. 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.
  2. 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.
  3. 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.
  4. 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.
  5. Mauricio S Antunes

    Department of Biology, Colorado State University, Fort Collins, United States
    Competing interests
    No competing interests declared.
  6. 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.
  7. Kevin J Morey

    Department of Biology, Colorado State University, Fort Collins, United States
    Competing interests
    No competing interests declared.
  8. Xavier Rios

    Department of Genetics, Harvard Medical School, Boston, United States
    Competing interests
    No competing interests declared.
  9. June I Medford

    Department of Biology, Colorado State University, Fort Collins, United States
    Competing interests
    No competing interests declared.
  10. 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.
  11. 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.
  12. 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

  1. Jeffery W Kelly, Scripps Research Institute, United States

Version history

  1. Received: August 6, 2015
  2. Accepted: December 17, 2015
  3. Accepted Manuscript published: December 29, 2015 (version 1)
  4. Accepted Manuscript updated: December 30, 2015 (version 2)
  5. Version of Record published: January 26, 2016 (version 3)

Copyright

© 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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  1. Justin Feng
  2. Benjamin W Jester
  3. Christine E Tinberg
  4. Daniel J Mandell
  5. Mauricio S Antunes
  6. Raj Chari
  7. Kevin J Morey
  8. Xavier Rios
  9. June I Medford
  10. George M Church
  11. Stanley Fields
  12. David Baker
(2015)
A general strategy to construct small molecule biosensors in eukaryotes
eLife 4:e10606.
https://doi.org/10.7554/eLife.10606

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https://doi.org/10.7554/eLife.10606

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