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Genome-scale CRISPRi screen identifies pcnB repression conferring improved physiology for overproduction of free fatty acids in Escherichia coli

ABSTRACT: Genome-scale CRISPRi screen identifies pcnB repression conferring improved physiology for overproduction of free fatty acids in Escherichia coli

PROVIDER: PRJNA1112435 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29060100_1.fastq.gz	Fastqsanger.gz
	SRR29060100_2.fastq.gz	Fastqsanger.gz
	SRR29060095_1.fastq.gz	Fastqsanger.gz
	SRR29060095_2.fastq.gz	Fastqsanger.gz
	SRR29060096_1.fastq.gz	Fastqsanger.gz

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Genome-scale CRISPRi screen identifies pcnB repression conferring improved physiology for overproduction of free fatty acids in Escherichia coli

Project description:Microbial physiology plays a pivotal role in construction of a superior microbial cell factory for efficient production of desired products. Here we identified pcnB repression through genome-scale CRISPRi modulation combining fluorescence-activated cell sorting (FACS) and next-generation sequencing (NGS), which confers improved physiology for free fatty acids (FFAs) overproduction in Escherichia coli. The repression of pcnB could improve the stability and abundance of the transcripts involved in proton-consuming system, conferring a global improvement on cell membrane, redox state, and energy level. These physiological advantages facilitated further identification of acrD repression enhancing FFAs efflux. The engineered strain pcnBi-acrDi-fadR+ achieved 35.1 g l−1 FFAs production in fed-batch fermentation, which is the maximum titer in E. coli reported to date. This study underscores the significance of hidden genetic determinants in microbial biosynthesis and sheds light on the role of microbial physiologies in boosting microbial biosynthesis.

2024-05-21 | GSE267710 | GEO

Genome-scale CRISPRi screen identifies pcnB repression conferring improved physiology for overproduction of free fatty acids in Escherichia coli II

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Genome-scale CRISPRi screen identifies pcnB repression conferring improved physiology for overproduction of free fatty acids in Escherichia coli II

Project description:Genome-scale CRISPRi screen identifies pcnB repression conferring improved physiology for overproduction of free fatty acids in Escherichia coli II

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Genome-scale CRISPRi screen identifies pcnB repression conferring overproduction of free fatty acids in Escherichia coli

Project description:In this study, we carried out genome-scale CRISPRi modulation combining fluorescence-activated cell sorting (FACS) and next-generation sequencing (NGS) to identify genetic determinants for FFAs overproduction in Escherichia coli. The pcnBi (AP) strain that repressed the expression of pcnB produced 2992 mg l FFAs, which was 4.0-fold of the CF (A) strain. To analyze the underlying mechanism of FFAs overproduction, the engineered strain pcnBi and the control strain CF were applied to the transcriptomic and proteomic analyses.

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Project description:In Mycobacterium tuberculosis, the efflux pump, EfpA, plays an essential but uncharacterised role in its physiology and antibiotic tolerance. In this study, an ATc-inducible CRISPR interference (CRISPRi) system was introduced into M. tuberculosis to knock-down the expression of efpA and determine any subsequent effects on the organism. As part of the study, microarray experiments were performed to determine any changes in gene expression caused by efpA repression M. tuberculosis by comparing the transcriptomic profile between ATc-induced efpA knock-down cultures to that of efpA knock-down cultures without ATc-induction.

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