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Title: Developing Riboswitch-Mediated Gene Regulatory Controls in Thermophilic Bacteria

Abstract

Thermophilic bacteria are attractive hosts to produce bio-based chemicals. While various genetic manipulations have been employed in the metabolic engineering of thermophiles, a robust means to regulate gene expression in these bacteria (~55 degrees C) is missing. Our bioinformatic search for various riboswitches in thermophilic bacteria revealed that major classes of riboswitches are present, suggesting riboswitches' regulatory roles in these bacteria. By building synthetic constructs incorporating natural and engineered purine riboswitch sequences originated from foreign species, we quantified respective riboswitches activities in repressing and up-regulating gene expression in Geobacillus thermoglucosidasius using a green fluorescence protein. The elicited regulatory response was ligand-concentration-dependent. We further demonstrated that riboswitch-mediated gene expression of adhE (responsible for ethanol production) in Clostridium thermocellum can modulate ethanol production, redirect metabolites, and control cell growth in the adhE knockout mutant. This work has made tunable gene expression feasible across different thermophiles for broad applications including biofuels production and gene-to-trait mapping.

Authors:
 [1];  [1];  [1];  [1];  [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE)
OSTI Identifier:
1507282
Alternate Identifier(s):
OSTI ID: 1505245
Report Number(s):
NREL/JA-2700-73607
Journal ID: ISSN 2161-5063
Grant/Contract Number:  
AC36-08GO28308; 06271601; AC36-08-GO28308
Resource Type:
Accepted Manuscript
Journal Name:
ACS Synthetic Biology
Additional Journal Information:
Journal Volume: 8; Journal Issue: 4; Journal ID: ISSN 2161-5063
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Clostridium thermocellum; gene regulation; inducible and repressible systems; riboswitch; thermophiles

Citation Formats

Marcano-Velazquez, Joan G., Lo, Jonathan, Nag, Ambarish, Maness, Pin-Ching, and Chou, Katherine J. Developing Riboswitch-Mediated Gene Regulatory Controls in Thermophilic Bacteria. United States: N. p., 2019. Web. doi:10.1021/acssynbio.8b00487.
Marcano-Velazquez, Joan G., Lo, Jonathan, Nag, Ambarish, Maness, Pin-Ching, & Chou, Katherine J. Developing Riboswitch-Mediated Gene Regulatory Controls in Thermophilic Bacteria. United States. doi:10.1021/acssynbio.8b00487.
Marcano-Velazquez, Joan G., Lo, Jonathan, Nag, Ambarish, Maness, Pin-Ching, and Chou, Katherine J. Mon . "Developing Riboswitch-Mediated Gene Regulatory Controls in Thermophilic Bacteria". United States. doi:10.1021/acssynbio.8b00487.
@article{osti_1507282,
title = {Developing Riboswitch-Mediated Gene Regulatory Controls in Thermophilic Bacteria},
author = {Marcano-Velazquez, Joan G. and Lo, Jonathan and Nag, Ambarish and Maness, Pin-Ching and Chou, Katherine J.},
abstractNote = {Thermophilic bacteria are attractive hosts to produce bio-based chemicals. While various genetic manipulations have been employed in the metabolic engineering of thermophiles, a robust means to regulate gene expression in these bacteria (~55 degrees C) is missing. Our bioinformatic search for various riboswitches in thermophilic bacteria revealed that major classes of riboswitches are present, suggesting riboswitches' regulatory roles in these bacteria. By building synthetic constructs incorporating natural and engineered purine riboswitch sequences originated from foreign species, we quantified respective riboswitches activities in repressing and up-regulating gene expression in Geobacillus thermoglucosidasius using a green fluorescence protein. The elicited regulatory response was ligand-concentration-dependent. We further demonstrated that riboswitch-mediated gene expression of adhE (responsible for ethanol production) in Clostridium thermocellum can modulate ethanol production, redirect metabolites, and control cell growth in the adhE knockout mutant. This work has made tunable gene expression feasible across different thermophiles for broad applications including biofuels production and gene-to-trait mapping.},
doi = {10.1021/acssynbio.8b00487},
journal = {ACS Synthetic Biology},
number = 4,
volume = 8,
place = {United States},
year = {2019},
month = {3}
}

Journal Article:
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This content will become publicly available on March 18, 2020
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