{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Ingle P"],"funding":["Swiss National Science Foundation","Medical Research Council","National Institute for Health Research (NIHR)","Biotechnology and Biological Sciences Research Council"],"pagination":["8123"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC6544763"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["9(1)"],"pubmed_abstract":["Understanding the molecular pathogenesis of Clostridioides difficile has relied on the use of ermB-based mutagens in erythromycin-sensitive strains. However, the repeated subcultures required to isolate sensitive variants can lead to the acquisition of ancillary mutations that affect phenotype, including virulence. CRISPR-Cas9 allows the direct selection of mutants, reducing the number of subcultures and thereby minimising the likelihood of acquiring additional mutations. Accordingly, CRISPR-Cas9 was used to sequentially remove from the C. difficile 630 reference strain (NCTC 13307) two ermB genes and pyrE. The genomes of the strains generated (630Δerm* and 630Δerm*ΔpyrE, respectively) contained no ancillary mutations compared to the NCTC 13307 parental strain, making these strains the pre"],"journal":["Scientific reports"],"pubmed_title":["Generation of a fully erythromycin-sensitive strain of Clostridioides difficile using a novel CRISPR-Cas9 genome editing system."],"pmcid":["PMC6544763"],"funding_grant_id":["BB/L01081X/1","147603","BB/L502030/1","BB/M012336/1","NIHR-INF-0155","BB/K00283X/1","BB/L013940/1","G0601176"],"pubmed_authors":["Ingle P","Cockayne A","Kuehne SA","Gu Y","Jiang W","Huang H","Groothuis D","Rowe P","Humphreys CM","Minton NP"],"additional_accession":[]},"is_claimable":false,"name":"Generation of a fully erythromycin-sensitive strain of Clostridioides difficile using a novel CRISPR-Cas9 genome editing system.","description":"Understanding the molecular pathogenesis of Clostridioides difficile has relied on the use of ermB-based mutagens in erythromycin-sensitive strains. However, the repeated subcultures required to isolate sensitive variants can lead to the acquisition of ancillary mutations that affect phenotype, including virulence. CRISPR-Cas9 allows the direct selection of mutants, reducing the number of subcultures and thereby minimising the likelihood of acquiring additional mutations. Accordingly, CRISPR-Cas9 was used to sequentially remove from the C. difficile 630 reference strain (NCTC 13307) two ermB genes and pyrE. The genomes of the strains generated (630Δerm* and 630Δerm*ΔpyrE, respectively) contained no ancillary mutations compared to the NCTC 13307 parental strain, making these strains the pre","dates":{"release":"2019-01-01T00:00:00Z","publication":"2019 May","modification":"2026-05-07T02:08:50.495Z","creation":"2019-07-24T07:07:55Z"},"accession":"S-EPMC6544763","cross_references":{"pubmed":["31148548"],"doi":["10.1038/s41598-019-44458-y"]}}