ArticleMetabolic engineering2023
Biosensor-assisted titratable CRISPRi high-throughput (BATCH) screening for over-production phenotypes.
Article in Metabolic engineering, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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18 citing papers in PubMed, 48 citations in OpenAlex.
- Advances in Zosteric Acid: Antifouling Properties and Green Biosynthesis Toward Food Safety Applications.Foods (Basel, Switzerland) · 2026Review
- Whole-cell and cell-free biosensor-driven metabolic engineering.Current opinion in biotechnology · 2025Review
- Characterizing and Engineering a Succinate-Responsive Biosensor System inACS synthetic biology · 2025Article
- Advancing lignocellulosic conversion though biosensor-enabled metabolic engineering.Green chemistry : an international journal and green chemistry resource : GC · 2025Review
- Engineering an Overflow-Responsive Regulation System for Balancing Cellular Redox and Optimizing Microbial Production.Biotechnology and bioengineering · 2025Article
- Metabolic engineering of Escherichia coli for enhanced production of p-coumaric acid via L-phenylalanine biosynthesis pathway.Bioprocess and biosystems engineering · 2025Article
- Genetically encoded biosensors for the circular plastics bioeconomy.Metabolic engineering communications · 2024Article
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- Investigating and Engineering an 1,2-Propanediol-Responsive Transcription Factor-Based Biosensor.ACS synthetic biology · 2024Article
- Tunable translation-level CRISPR interference by dCas13 and engineered gRNA in bacteria.Nature communications · 2024Article
- Biosensor-assisted CRISPRi high-throughput screening to identify genetic targets inSynthetic and systems biotechnology · 2024Article
- The pAblo·pCasso self-curing vector toolset for unconstrained cytidine and adenine base-editing in Gram-negative bacteria.Nucleic acids research · 2024Article
- Application of functional genomics for domestication of novel non-model microbes.Journal of industrial microbiology & biotechnology · 2024Review
- The expanded CRISPR toolbox for constructing microbial cell factories.Trends in biotechnology · 2024Review
- Establishing Tunable Genetic Logic Gates with Versatile Dynamic Performance by Varying Regulatory Parameters.ACS synthetic biology · 2023Article
- Genome-scale CRISPRi screening: A powerful tool in engineering microbiology.Engineering microbiology · 2023Review
- Dynamic Metabolic Control: From the Perspective of Regulation Logic.Synthetic biology and engineering · 2023Article
- Transcription factor-based biosensors for screening and dynamic regulation.Frontiers in bioengineering and biotechnology · 2023Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
With rapid advances in the development of metabolic pathways and synthetic biology toolkits, a persisting challenge in microbial bioproduction is how to optimally rewire metabolic fluxes and accelerate the concomitant high-throughput phenotype screening. Here we developed a biosensor-assisted titratable CRISPRi high-throughput (BATCH) screening approach that combines a titratable mismatch CRISPR interference and a biosensor mediated screening for high-production phenotypes in Escherichia coli. We first developed a programmable mismatch CRISPRi that could afford multiple levels of interference efficacy with a one-pot sgRNA pool (a total of 16 variants for each target gene) harboring two consecutive random mismatches in the seed region of sgRNA spacers. The mismatch CRISPRi was demonstrated to enable almost a full range of gene knockdown when targeting different positions on genes. As a proof-of-principle demonstration of the BATCH screening system, we designed doubly mismatched sgRNA pools targeting 20 relevant genes in E. coli and optimized a PadR-based p-coumaric acid biosensor with broad dynamic range for the eGFP fluorescence guided high-production screening. Using sgRNA variants for the combinatorial knockdown of pfkA and ptsI, the p-coumaric acid titer was increased by 40.6% to o 1308.6 mg/l from glycerol in shake flasks. To further demonstrate the general applicability of the BATCH screening system, we recruited a HpdR-based butyrate biosensor that facilitated the screening of E. coli strains achieving 19.0% and 25.2% increase of butyrate titer in shake flasks with sgRNA variants targeting sucA and ldhA, respectively. This work reported the establishment of a plug-and-play approach that enables multilevel modulation of metabolic fluxes and high-throughput screening of high-production phenotypes.
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