ArticleScientific reports2017
Using CRISPR-Cas9-mediated genome editing to generate C. difficile mutants defective in selenoproteins synthesis.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 68 papers.
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Who cites it
68 citing papers in PubMed.
- Transferable genetic toolsets for nonmodel gut Clostridia enable in vivo reversible control of metabolite production.Nature biotechnology · 2026Article
- A set of genetic tools for use inMicrobiology (Reading, England) · 2026Article
- CRISPR-AsCas12a and dAsCas12a-Mediated Gene Knockout and Knockdown in Clostridioides difficile.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Nanoluciferase-Based Reporters for Clostridioides difficile.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Visualizing C. difficile During Murine Infection.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Generating Targeted Mutants Using a Riboswitch to Control Plasmid Behavior During Allelic Exchange Mutagenesis.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Flagellar switch inverted repeats impact heterogeneity in flagellar gene expression and thus C. difficile RT027/MLST1 virulence.Cell reports · 2025Article
- A thiouracil desulfurase protects Clostridioides difficile RNA from 4-thiouracil incorporation, providing a competitive advantage in the gut.Cell host & microbe · 2025Article
- Identification of two glycosyltransferases required for synthesis of membrane glycolipids inmBio · 2025Article
- Deletion of atypical type II restriction genes in Clostridium cellulovorans using a Cas9-based gene editing system.Applied microbiology and biotechnology · 2025Article
- Identification of two glycosyltransferases required for synthesis of membrane glycolipids inbioRxiv : the preprint server for biology · 2025Article
- Article
- The Impact of YabG Mutations on Clostridioides difficile Spore Germination and Processing of Spore Substrates.Molecular microbiology · 2024Article
- Adaptation mechanisms of Clostridioides difficile to auranofin and its impact on human gut microbiota.NPJ biofilms and microbiomes · 2024Article
- Identification of a family of peptidoglycan transpeptidases reveals thatProceedings of the National Academy of Sciences of the United States of America · 2024Article
- Genetic manipulations of nonmodel gut microbes.iMeta · 2024Article
- Theophylline-based control of repA on a Clostridioides difficile plasmid for use in allelic exchange.Anaerobe · 2024Article
- The impact of YabG mutations onbioRxiv : the preprint server for biology · 2024Article
- Revealing roles of S-layer protein (SlpA) inMicrobiology spectrum · 2024Article
- Identification ofJournal of bacteriology · 2024Article
8 more citing papers are in PubMed but not listed here.
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5 authors.
Funding
Abstract
Clostridium difficile is a significant concern as a nosocomial pathogen, and genetic tools are important when analyzing the physiology of such organisms so that the underlying physiology/pathogenesis of the organisms can be studied. Here, we used TargeTron to investigate the role of selenoproteins in C. difficile Stickland metabolism and found that a TargeTron insertion into selD, encoding the selenophosphate synthetase that is essential for the specific incorporation of selenium into selenoproteins, results in a significant growth defect and a global loss of selenium incorporation. However, because of potential polar effects of the TargeTron insertion, we developed a CRISPR-Cas9 mutagenesis system for C. difficile. This system rapidly and efficiently introduces site-specific mutations into the C. difficile genome (20-50% mutation frequency). The selD CRISPR deletion mutant had a growth defect in protein-rich medium and mimicked the phenotype of a generated TargeTron selD mutation. Our findings suggest that Stickland metabolism could be a target for future antibiotic therapies and that the CRISPR-Cas9 system can introduce rapid and efficient modifications into the C. difficile genome.
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