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

Carrie Eckert, Rongming Liu, Liya Liang, Alaksh Choudhury, Andrew Garst, Eun Oh, James Winkler, Ryan Gill

Research output: Contribution to journalArticlepeer-review

31 Scopus Citations


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.
Original languageAmerican English
Pages (from-to)50-58
Number of pages9
JournalMetabolic Engineering
StatePublished - 2019

NREL Publication Number

  • NREL/JA-2700-72224


  • combinatorial programming
  • gene expression
  • global regulatory networks
  • multiplex navigation


Dive into the research topics of 'Multiplex Navigation of Global Regulatory Networks (MINR) in Yeast for Improved Ethanol Tolerance and Production'. Together they form a unique fingerprint.

Cite this