Genome‐Wide CRISPRi Screening of Key Genes for Recombinant Protein Expression in Bacillus Subtilis
Abstract: Bacillus subtilis is an industrially important microorganism that is often used as a microbial cell factory for the production of recombinant proteins due to its food safety, rapid growth, and powerful secretory capacity. However, the lack of data on functional genes related to recombinant protein production has hindered the further development of B. subtilis cell factories. Here, a strategy combining genome‐wide CRISPRi screening and targeted CRISPRa activation to enhance recombinant protein expression is proposed. First, a CRISPRi library covering a total of 4225 coding genes (99.7%) in the B. subtilis genome and built the corresponding high‐throughput screening methods is constructed. Twelve key genes for recombinant protein expression are identified, including targets without relevant functional annotations. Meanwhile, the transcription of recombinant protein genes by CRISPRa is up‐regulated. These screened or selected genes can be easily applied to metabolic engineering by constructing sgRNA arrays. The relationship between differential pathways and recombinant protein expression in engineered strains by transcriptome analysis is also revealed. High‐density fermentation and generalisability validation results prove the reliability of the strategy. This method can be extended to other industrial hosts to support functional gene annotation and the design of novel cell factories.
- Location
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Deutsche Nationalbibliothek Frankfurt am Main
- Extent
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Online-Ressource
- Language
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Englisch
- Bibliographic citation
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Genome‐Wide CRISPRi Screening of Key Genes for Recombinant Protein Expression in Bacillus Subtilis ; day:01 ; month:07 ; year:2024 ; extent:13
Advanced science ; (01.07.2024) (gesamt 13)
- Creator
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Zhu, Xuyang
Luo, Hui
Yu, Xinrui
Lv, Huihui
Su, Lingqia
Zhang, Kang
Wu, Jing
- DOI
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10.1002/advs.202404313
- URN
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urn:nbn:de:101:1-2407021421395.034373530351
- Rights
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Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Last update
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14.08.2025, 10:50 AM CEST
Data provider
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Associated
- Zhu, Xuyang
- Luo, Hui
- Yu, Xinrui
- Lv, Huihui
- Su, Lingqia
- Zhang, Kang
- Wu, Jing
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