公开(公告)号：US20230212626A1

公开(公告)日：2023-07-06

申请号：US18087169

申请日：2022-12-22

Applicant: The Trustees of Princeton University

Inventor： Jose L. Avalos ,  Jared E. Toettcher ,  Evan M. Zhao ,  Makoto A. Lalwani

IPC: C12P17/06 ,  C12N1/16 ,  C12N15/63 ,  C12N15/81 ,  C12P7/56 ,  C12N9/10 ,  C12N9/88 ,  C12N9/04

CPC classification number: C12P17/06 ,  C12N1/16 ,  C12N15/635 ,  C12N15/81 ,  C12P7/56 ,  C12N9/1022 ,  C12Y202/01006 ,  C12N9/88 ,  C12Y401/01005 ,  C12Y403/01023 ,  C12Y403/01024 ,  C12N9/0006 ,  C12Y101/01027 ,  C12N2529/10 ,  C12N2800/102 ,  C12N2830/002

Abstract: Disclosed is a technique for constructing optogenetic amplifier and inverter circuits utilizing transcriptional activator/repressor pairs, in which expression of the transcriptional activator or repressor, respectively, is controlled by light-controlled transcription factors. This system is demonstrated utilizing the quinic acid regulon system from Neurospora crassa, or Q System, a transcriptional activator/repressor system. This is also demonstrated utilizing the galactose regulon from Saccharomyces cerevisiae, or GAL System. Such optogenetic amplifier circuits enable multi-phase microbial fermentations, in which different light schedules are applied in each phase to dynamically control different metabolic pathways for the production of proteins, fuels or chemicals. The orthogonal nature of the Q and GAL systems enable the co-expression of amplifier and inverter circuits to simultaneously amplify and invert the response of light-controlled transcriptional controls over different sets of genes in the same cell.

公开(公告)号：US20210062165A1

公开(公告)日：2021-03-04

申请号：US17003659

申请日：2020-08-26

Applicant: The Trustees of Princeton University

Inventor： Jose L. Avalos ,  Jared E. Toettcher ,  Clifford P. Brangwynne ,  Evan M. Zhao ,  Maxwell Z. Wilson

IPC: C12N9/02 ,  C12N13/00 ,  C12N15/81

Abstract: Provided herein is a system and method of optogenetically inducibly clustering metabolic enzymes for the production of chemicals using cell factories. More particularly, the described inducible protein clustering approach clusters metabolic enzymes by, e.g., a change in illumination conditions (either a switch from dark to light or from light to dark). Performing this clustering leads to an increase in the production of metabolites by the clustered enzymes. In some embodiments, a light-sensitive domain may be replaced with any inducible domain.

公开(公告)号：US20200291075A1

公开(公告)日：2020-09-17

申请号：US16889690

申请日：2020-06-01

Applicant: The Trustees of Princeton University

Inventor： Jared Toettcher ,  Jose Avalos ,  Maxwell Wilson ,  Alexander Goglia ,  Evan M. Zhao ,  Agnieszka Gil ,  Cesar Carrasco-Lopez

IPC: C07K14/415 ,  C07K1/16 ,  C07K1/36 ,  C07K19/00

Abstract: Provided herein is an isolated fusion protein comprising a light-responsive domain and a heterologous peptide component. Exposure of the fusion protein to light induces a conformational change in the fusion protein that alters an activity of the fusion protein and the activity is a binding activity selected from an in vitro binding activity, an in vivo extracellular binding activity and an in vivo intracellular binding activity. Also provided are methods of using and identifying the fusion proteins.

公开(公告)号：US11859223B2

公开(公告)日：2024-01-02

申请号：US16091624

申请日：2017-04-07

Applicant: The Trustees of Princeton University

Inventor： Jose L. Avalos ,  Jared E. Toettcher ,  Evan M. Zhao

IPC: C12P7/56 ,  C12N9/04 ,  C12N9/88 ,  C12N9/10 ,  C12N13/00 ,  C12P7/16 ,  C12N15/81 ,  C12N15/63

CPC classification number: C12N9/88 ,  C12N9/0006 ,  C12N9/1022 ,  C12N13/00 ,  C12N15/63 ,  C12N15/635 ,  C12N15/81 ,  C12P7/16 ,  C12P7/56 ,  C12Y101/01027 ,  C12Y202/01006 ,  C12Y401/01001

Abstract: A system and method for controlling metabolic enzymes or pathways in cells to produce a chemical above the levels of a wild-type strain is disclosed. The system utilizes cells, including yeasts, bacteria, and molds, having at least two genes capable of being controlled bi-directionally with light, where one gene is turned from off to on when exposed to light and another gene is turned from on to off when exposed to light, the two genes reversing when the light is turned off. Cells may utilize any number of sequences that benefit chemical production, including sequences that: encode for constitutive transcription of light-activated transcription factor fusions; encode for a metabolic enzyme; encode for a repressor; induce expression of metabolic enzymes; and an endogenous or exogenous activator expressed by a constitutive promoter, inducible promoter, or gene circuit. These systems may be coupled to biosensors or protein cascade systems, enabling the monitoring or automation of the fermentation process to optimize production of a desired product. These systems may also allow for optimization and periodic operation of a bioreactor using light pulses.

公开(公告)号：US11746131B2

公开(公告)日：2023-09-05

申请号：US16889690

申请日：2020-06-01

Applicant: The Trustees of Princeton University

Inventor： Jared Toettcher ,  Jose Avalos ,  Maxwell Wilson ,  Alexander Goglia ,  Evan M. Zhao ,  Agnieszka Gil ,  Cesar Carrasco-Lopez

IPC: C07K14/415 ,  C07K19/00 ,  C07K1/16 ,  C07K1/36

CPC classification number: C07K14/415 ,  C07K1/16 ,  C07K1/36 ,  C07K19/00 ,  C07K2319/80

Abstract: Provided herein is an isolated fusion protein comprising a light-responsive domain and a heterologous peptide component. Exposure of the fusion protein to light induces a conformational change in the fusion protein that alters an activity of the fusion protein and the activity is a binding activity selected from an in vitro binding activity, an in vivo extracellular binding activity and an in vivo intracellular binding activity. Also provided are methods of using and identifying the fusion proteins.