ArticleAnaerobe2024
Theophylline-based control of repA on a Clostridioides difficile plasmid for use in allelic exchange.
Article in Anaerobe, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
What it found
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Who cites it
9 citing papers in PubMed, 12 citations in OpenAlex.
- Phase variable colony morphotypes ofInfection and immunity · 2026Article
- Phase variable colony morphotypes ofbioRxiv : the preprint server for biology · 2026Article
- Phenotypic analysis of variousApplied and environmental microbiology · 2025Article
- Article
- Development of a conditional plasmid for gene deletion in non-modelApplied and environmental microbiology · 2025Article
- The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates.Molecular microbiology · 2024Article
- The small acid-soluble proteins of Clostridioides difficile regulate sporulation in a SpoIVB2-dependent manner.PLoS pathogens · 2024Article
- The small acid-soluble proteins ofbioRxiv : the preprint server for biology · 2024Article
- The impact of YabG mutations onbioRxiv : the preprint server for biology · 2024Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Historically, mutagenesis in the non-model enteropathogenic bacterium Clostridioides difficile has been challenging. Developing a versatile and reliable method of generating targeted mutations in C. difficile is important to further our understanding of its pathogenesis. Some of the most common targeted mutagenesis systems rely on allelic exchange mediated by either uracil auxotrophy combined with a toxic uracil precursor, a toxin/anti-toxin system, group II introns, or CRISPR/Cas mutagenesis. However, each of these methods suffers from its own issues. Here, we develop and test an allelic exchange strategy which better facilitates screening for integration and selecting for excision than previous systems. This is achieved by controlling plasmid replication with a theophylline-dependent riboswitch cloned upstream of repA, the gene whose product controls plasmid replication. This allows efficient mutant generation, can be performed in a wild-type strain of C. difficile, does not have the off-target effects inherent to group II introns, and alleviates the problem of testing multiple gRNA targets in CRISPR mutagenesis.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.