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This content will become publicly available on December 29, 2015

Title: 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. As a result, this work provides a general methodology to develop biosensors for a broad range of molecules in eukaryotes.
Authors:
 [1] ;  [2] ;  [2] ;  [3] ;  [4] ;  [1] ;  [4] ;  [1] ;  [4] ;  [5] ;  [2] ;  [2]
  1. Harvard Medical School, Boston, MA (United States)
  2. Univ. of Washington, Seattle, WA (United States)
  3. Harvard Medical School, Boston, MA (United States); Harvard Univ., Boston, MA (United States)
  4. Colorado State Univ., Fort Collins, CO (United States)
  5. Harvard Medical School, Boston, MA (United States); Harvard Univ., Boston, (United States)
Publication Date:
OSTI Identifier:
1235744
Grant/Contract Number:
FG02-02ER63445; FG02- 02ER63445
Type:
Published Article
Journal Name:
eLife
Additional Journal Information:
Journal Volume: 4; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Research Org:
Harvard Univ., Cambridge, MA (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES