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Title: Multiplex Navigation of Global Regulatory Networks (MINR) in yeast for Improved Ethanol Tolerance and Production

Abstract

Multiplex navigation of global regulatory networks (MINR) is an approach for combinatorially reprogramming gene expression to manipulate complex phenotypes. We designed, constructed, and mapped MINR libraries containing 43,020 specific mutations in 25 regulatory genes expected to perturb the yeast regulatory network. We selected growth competition experiments for library mutants conferring increased ethanol and/or glucose tolerance. We identified specific mutants that not only possessed improved ethanol and/or glucose tolerance but also produced ethanol at concentrations up to 2-fold higher than those produced by the wild-type strain. We further determined that mutations increasing ethanol tolerance were transferable to a diploid industrial yeast strain. The facile construction and mapping of 43,020 designer regulatory mutations provide a roadmap for how to access and engineer complex phenotypes in future synthetic biology and broader efforts.

Authors:
 [1];  [1];  [1];  [1];  [2];  [1];  [1];  [1]
  1. Univ. of Colorado, Boulder, CO (United States)
  2. Univ. of Colorado, Boulder, CO (United States); 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), Bioenergy Technologies Office (EE-3B)
OSTI Identifier:
1465647
Report Number(s):
NREL/JA-2700-72224
Journal ID: ISSN 1096-7176
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Accepted Manuscript
Journal Name:
Metabolic Engineering
Additional Journal Information:
Journal Volume: 51; Journal ID: ISSN 1096-7176
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; multiplex navigation; global regulatory networks; combinatorial programming; gene expression

Citation Formats

Liu, Rongming, Liang, Liya, Choudhury, Alaksh, Garst, Andrew D., Eckert, Carrie A., Oh, Eun Joong, Winkler, James, and Gill, Ryan T. Multiplex Navigation of Global Regulatory Networks (MINR) in yeast for Improved Ethanol Tolerance and Production. United States: N. p., 2018. Web. doi:10.1016/j.ymben.2018.07.007.
Liu, Rongming, Liang, Liya, Choudhury, Alaksh, Garst, Andrew D., Eckert, Carrie A., Oh, Eun Joong, Winkler, James, & Gill, Ryan T. Multiplex Navigation of Global Regulatory Networks (MINR) in yeast for Improved Ethanol Tolerance and Production. United States. doi:10.1016/j.ymben.2018.07.007.
Liu, Rongming, Liang, Liya, Choudhury, Alaksh, Garst, Andrew D., Eckert, Carrie A., Oh, Eun Joong, Winkler, James, and Gill, Ryan T. Tue . "Multiplex Navigation of Global Regulatory Networks (MINR) in yeast for Improved Ethanol Tolerance and Production". United States. doi:10.1016/j.ymben.2018.07.007. https://www.osti.gov/servlets/purl/1465647.
@article{osti_1465647,
title = {Multiplex Navigation of Global Regulatory Networks (MINR) in yeast for Improved Ethanol Tolerance and Production},
author = {Liu, Rongming and Liang, Liya and Choudhury, Alaksh and Garst, Andrew D. and Eckert, Carrie A. and Oh, Eun Joong and Winkler, James and Gill, Ryan T.},
abstractNote = {Multiplex navigation of global regulatory networks (MINR) is an approach for combinatorially reprogramming gene expression to manipulate complex phenotypes. We designed, constructed, and mapped MINR libraries containing 43,020 specific mutations in 25 regulatory genes expected to perturb the yeast regulatory network. We selected growth competition experiments for library mutants conferring increased ethanol and/or glucose tolerance. We identified specific mutants that not only possessed improved ethanol and/or glucose tolerance but also produced ethanol at concentrations up to 2-fold higher than those produced by the wild-type strain. We further determined that mutations increasing ethanol tolerance were transferable to a diploid industrial yeast strain. The facile construction and mapping of 43,020 designer regulatory mutations provide a roadmap for how to access and engineer complex phenotypes in future synthetic biology and broader efforts.},
doi = {10.1016/j.ymben.2018.07.007},
journal = {Metabolic Engineering},
number = ,
volume = 51,
place = {United States},
year = {2018},
month = {7}
}

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