Figure 1.

Improving bacterial tolerance to biofuels using CRISPR gene expression perturbation. (A) CRISPR perturbations of gene expression (both inhibition and activation) were designed for 31 E. coli genes and expressed using a two-plasmid system. (B) Strains used in this study. Whether CRISPR constructs were used to inhibit or activate gene expression are notated by -i or -a in the strain name respectively. (C) These perturbations were designed to disrupt expression of a variety of genes related to bacterial metabolism, redox, transport, various DNA and RNA processes, and motility. (D) Strains harboring these individual perturbations were exposed to biofuels (n-butanol and n-hexane) over multiple days, with the characterization of both short-term and long-term growth impacts.