ArticleBiotechnology and bioengineering2020
CRISPR interference of nucleotide biosynthesis improves production of a single-domain antibody in Escherichia coli.
Article in Biotechnology and bioengineering, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 20 citations in OpenAlex.
- The landscape of antibody production systems: recombinant expression for research, diagnostics and therapy.Frontiers in bioengineering and biotechnology · 2026Review
- Exploring the Dynamic Culture System of biofilm-forming Bifidobacterium Adolescentis BL-8: from System establishment, Formation Mechanisms Analysis To Application in freeze-dried Probiotic Powder.Current microbiology · 2025Article
- Expanding the CRISPR/Cas toolkit: applications in proteomics and theranostics.Frontiers in bioengineering and biotechnology · 2025Review
- Discovery of nanobodies: a comprehensive review of their applications and potential over the past five years.Journal of nanobiotechnology · 2024Review
- Engineering activatable promoters for scalable and multi-input CRISPRa/i circuits.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- A serum NMR metabolomic analysis of the Corynebacterium pseudotuberculosis infection in goats.Applied microbiology and biotechnology · 2023Article
- Large-Scale Production of Anti-RNase A VHH Expressed inCurrent issues in molecular biology · 2023Article
- Isobutanol production by combinedMetabolic engineering communications · 2022Article
- Decoupling Growth and Production by Removing the Origin of Replication from a Bacterial Chromosome.ACS synthetic biology · 2022Article
- Review
- CRISPR interference of nucleotide biosynthesis improves production of a single-domain antibody in Escherichia coli.Biotechnology and bioengineering · 2020Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Growth decoupling can be used to optimize the production of biochemicals and proteins in cell factories. Inhibition of excess biomass formation allows for carbon to be utilized efficiently for product formation instead of growth, resulting in increased product yields and titers. Here, we used CRISPR interference to increase the production of a single-domain antibody (sdAb) by inhibiting growth during production. First, we screened 21 sgRNA targets in the purine and pyrimidine biosynthesis pathways and found that the repression of 11 pathway genes led to the increased green fluorescent protein production and decreased growth. The sgRNA targets pyrF, pyrG, and cmk were selected and further used to improve the production of two versions of an expression-optimized sdAb. Proteomics analysis of the sdAb-producing pyrF, pyrG, and cmk growth decoupling strains showed significantly decreased RpoS levels and an increase of ribosome-associated proteins, indicating that the growth decoupling strains do not enter stationary phase and maintain their capacity for protein synthesis upon growth inhibition. Finally, sdAb production was scaled up to shake-flask fermentation where the product yield was improved 2.6-fold compared to the control strain with no sgRNA target sequence. An sdAb content of 14.6% was reached in the best-performing pyrG growth decoupling strain.
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