BER launches Environmental System Science Program. Visit our new website under construction!

U.S. Department of Energy Office of Biological and Environmental Research

BER Research Highlights


Redirecting Metabolic Flux in E. Coli via Combinatorial CRISPRi-Mediated Repression for Isopentenol Production
Published: January 25, 2019
Posted: August 02, 2021

Objective

Use CRISPR interference (CRISPRi) to knock down endogenous genes in competing pathways to redirect metabolic flux toward target metabolite.

Approach

CRISPRi-mediated multiplex repression system to silence transcription of several endogenous genes to increase precursor availability in a heterologous isopentenol biosynthesis pathway.

Results/Impacts

  • Designed a single-gRNA library with 15 individual knockdown targets, where 3 gRNA cassettes targeting genes asnA, prpE, and gldA increased isopentenol titer by 18% to 24%.
  • Combined the three single-gRNA cassettes into a two- or three-gRNA array and observed up to 98% enhancement in production by fine-tuning the repression level through titrating dCas9 expression.
  • Strategy shows that multiplex combinatorial knockdown of competing genes using CRISPRi can increase production of the target metabolite.

Tian, T. et al. “Redirecting metabolic flux via combinatorial multiplex CRISPRi-mediated repression for isopentenol production in Escherichia coli.” ACS Synthetic Biology 8(2), 391–402 (2019). [DOI:10.1021/acssynbio.8b00429]

Topic Areas:

  • Research Area: Genomic Analysis and Systems Biology
  • Research Area: Microbes and Communities
  • Research Area: Sustainable Biofuels and Bioproducts
  • Research Area: DOE Bioenergy Research Centers (BRC)

Division: SC-33.2 Biological Systems Science Division, BER

 

BER supports basic research and scientific user facilities to advance DOE missions in energy and environment. More about BER

Recent Highlights

Jan 11, 2022
No Honor Among Copper Thieves
Findings provide a novel means to manipulate methanotrophs for a variety of environmental and in [more...]

Dec 06, 2021
New Genome Editing Tools Can Edit Within Microbial Communities
Two new technologies allow scientists to edit specific species and genes within complex laborato [more...]

Oct 27, 2021
Fungal Recyclers: Fungi Reuse Fire-Altered Organic Matter
Degrading pyrogenic (fire-affected) organic matter is an important ecosystem function of fungi i [more...]

Oct 19, 2021
Microbes Offer a Glimpse into the Future of Climate Change
Scientists identify key features in microbes that predict how warming affects carbon dioxide emi [more...]

Aug 25, 2021
Assessing the Production Cost and Carbon Footprint of a Promising Aviation Biofuel
Biomass-derived DMCO has the potential to serve as a low-carbon, high-performance jet fuel blend [more...]

List all highlights (possible long download time)