Small molecule biosensors
Abstract
Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Biosensors are produced based on a ligand-binding domain (LBD) using a method that, in principle, can be applied for 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. The power of this method is illustrated by developing biosensors for digoxin and progesterone. Addition of ligand to cells expressing a biosensor activates transcription in yeast, mammalian cells and plants, with a dynamic range of up to about 100-fold or more. The biosensors are used 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.
- Inventors:
- Issue Date:
- Research Org.:
- Harvard Univ., Cambridge, MA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 2221943
- Patent Number(s):
- 11693017
- Application Number:
- 15/760,352
- Assignee:
- President and Fellows of Harvard College (Cambridge, MA)
- DOE Contract Number:
- FG02-02ER63445
- Resource Type:
- Patent
- Resource Relation:
- Patent File Date: 01/12/2016
- Country of Publication:
- United States
- Language:
- English
Citation Formats
Mandell, Daniel J., Feng, Justin, Rios Villanueva, Xavier, and Chari, Rajagopal. Small molecule biosensors. United States: N. p., 2023.
Web.
Mandell, Daniel J., Feng, Justin, Rios Villanueva, Xavier, & Chari, Rajagopal. Small molecule biosensors. United States.
Mandell, Daniel J., Feng, Justin, Rios Villanueva, Xavier, and Chari, Rajagopal. Tue .
"Small molecule biosensors". United States. https://www.osti.gov/servlets/purl/2221943.
@article{osti_2221943,
title = {Small molecule biosensors},
author = {Mandell, Daniel J. and Feng, Justin and Rios Villanueva, Xavier and Chari, Rajagopal},
abstractNote = {Biosensors for small molecules can be used in applications that range from metabolic engineering to orthogonal control of transcription. Biosensors are produced based on a ligand-binding domain (LBD) using a method that, in principle, can be applied for 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. The power of this method is illustrated by developing biosensors for digoxin and progesterone. Addition of ligand to cells expressing a biosensor activates transcription in yeast, mammalian cells and plants, with a dynamic range of up to about 100-fold or more. The biosensors are used 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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2023},
month = {7}
}
Works referenced in this record:
CRISPR/CAS Systems for Genomic Modification and Gene Modulation
patent-application, September 2014
- Wu, Fangting
- US Patent Application 14/216655; 20140273226
High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells
journal, June 2013
- Fu, Yanfang; Foden, Jennifer A.; Khayter, Cyd
- Nature Biotechnology, Vol. 31, Issue 9
Structure and functions of powerful transactivators: VP16, MyoD and FoxA
journal, January 2010
- Hirai, Hiroyuki; Tani, Tetsuya; Kikyo, Nobuaki
- The International Journal of Developmental Biology, Vol. 54, Issue 11-12
Computational design of ligand-binding proteins with high affinity and selectivity
journal, September 2013
- Tinberg, Christine E.; Khare, Sagar D.; Dou, Jiayi
- Nature, Vol. 501, Issue 7466, p. 212-216
A Rapid, Reversible, and Tunable Method to Regulate Protein Function in Living Cells Using Synthetic Small Molecules
journal, September 2006
- Banaszynski, Laura A.; Chen, Ling-chun; Maynard-Smith, Lystranne A.
- Cell, Vol. 126, Issue 5
Broadly Applicable Methodology for the Rapid and Dosable Small Molecule-Mediated Regulation of Transcription Factors in Human Cells
journal, May 2013
- Shoulders, Matthew D.; Ryno, Lisa M.; Cooley, Christina B.
- Journal of the American Chemical Society, Vol. 135, Issue 22
Structure and function of the Zn(II) binding site within the DNA-binding domain of the GAL4 transcription factor.
journal, May 1989
- Pan, T.; Coleman, J. E.
- Proceedings of the National Academy of Sciences, Vol. 86, Issue 9