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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and CRP-E182D Transcriptomes

ABSTRACT: Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and CRP-E182D Transcriptomes

PROVIDER: PRJNA819865 | ENA |

REPOSITORIES: ENA

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Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and CRP-E182D Transcriptomes

Project description:Strain engineering for industrial production requires the improvement of tolerance to multiple unfavorable conditions. Here, we report using global regulator libraries based on the CRISPR-enabled trackable genome engineering (CREATE) method to engineer tolerance against multiple inhibitors in Escherichia coli. Deep mutagenesis libraries were rationally designed, constructed, and screened to target 34,340 mutations across 23 global regulators. A total of 69 specific mutations that respectively conferred tolerance to acetate, NaCl, furfural, and high temperature were isolated, confirmed, and evaluated. Among them, 32 novel reconstructed mutations exhibited better tolerance to the corresponding inhibitors, and the most dramatic mutation CRP-E182D conferred high cross-tolerance to acetate, NaCl and isobutanol. To further investigate the effects of this mutation in the CRP on acetate tolerance, whole-transcriptome sequencing (RNA-Seq) analysis was performed.

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