ArticleNucleic acids research2025
A CRISPR-SpCas9M-reporting system for efficient and rapid genome editing in Caulobacter crescentus.
Article in Nucleic acids research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.
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Who cites it
3 citing papers in PubMed.
- RECC: A Red/ET-CRISPR/Cas9-based system enabling genome mining of marineSynthetic and systems biotechnology · 2026Article
- Engineering genetic elements for microbial protein expression systems: Advances, challenges, applications, and prospects.Synthetic and systems biotechnology · 2026Review
- A Practical CRISPR-Based Method for Rapid Genome Editing inBio-protocol · 2025Article
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Authors and funding
21 authors.
Funding
Abstract
As members of the α-proteobacteria group, Caulobacter crescentus and its relatives are wildly studied for their unique asymmetric life cycle and versatile applications in industry, agriculture, and biomedicine. However, genetic manipulation in these bacteria remains challenging, typically requiring time-consuming and labor-intensive procedures. Here, we report a practical CRISPR-SpCas9M-reporting system that overcomes the limitations of SpCas9 expression and CRISPR escape, enabling efficient, markerless, and rapid genome editing in C. crescentus. Two genes encoding for a pair of scaffold proteins were knocked out individually or iteratively, demonstrating their direct involvements in cellular signaling asymmetry. Key components, including the Cas protein, Cas inducer, sgRNA, homologous arms, and reporter, were systematically analyzed and optimized in the system, finally achieving the apparent editing efficiency up to 80% in C. crescentus. Furthermore, we applied the CRISPR-SpCas9M-reporting system to two C. crescentus relatives, Agrobacterium fabrum and Sinorhizobium meliloti, establishing it as an efficient and general editing strategy. We anticipate that this system could be applied to other CRISPR-Cas-recalcitrant organisms, accelerating both basic and applied research in α-proteobacteria.
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Registered trials
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