The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
Jul. 04, 2023

Filed:

Jan. 12, 2016
Applicant:

President and Fellows of Harvard College, Cambridge, MA (US);

Inventors:

Daniel J. Mandell, Brookline, MA (US);

Justin Feng, Cambridge, MA (US);

Xavier Rios Villanueva, Boston, MA (US);

Rajagopal Chari, Brighton, MA (US);

Assignee:
Attorney:
Primary Examiner:
Int. Cl.
CPC ...
G01N 33/74 (2006.01); C07K 14/00 (2006.01); C12N 15/62 (2006.01); C12N 15/63 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/90 (2006.01);
U.S. Cl.
CPC ...
G01N 33/743 (2013.01); C07K 14/00 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/62 (2013.01); C12N 15/63 (2013.01); C12N 15/902 (2013.01); C07K 2319/00 (2013.01); C12N 2310/20 (2017.05); C12N 2320/50 (2013.01); C12N 2800/22 (2013.01); C12N 2800/80 (2013.01);
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.


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