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Cas9

  • Open Access
    Combined and Distinct Roles of Agr Proteins in <span class="named-content genus-species" id="named-content-1">Clostridioides difficile</span> 630 Sporulation, Motility, and Toxin Production
    Research Article | Host-Microbe Biology
    Combined and Distinct Roles of Agr Proteins in Clostridioides difficile 630 Sporulation, Motility, and Toxin Production

    C. difficile is a spore-forming, toxigenic, anaerobic bacterium that causes severe gastrointestinal illness. Understanding the ways in which C. difficile senses growth conditions to regulate toxin expression and sporulation is essential to advancing our understanding of this pathogen.

    ...
    Ummey Khalecha Bintha Ahmed, Tyler M. Shadid, Jason L. Larabee, Jimmy D. Ballard
  • Open Access
    A CRISPR Interference System for Efficient and Rapid Gene Knockdown in <span class="named-content genus-species" id="named-content-1">Caulobacter crescentus</span>
    Research Article | Molecular Biology and Physiology
    A CRISPR Interference System for Efficient and Rapid Gene Knockdown in Caulobacter crescentus

    Caulobacter crescentus is a major model organism for understanding cell cycle regulation and cellular asymmetry. The current genetic tools for deleting or silencing the expression of individual genes, particularly those essential for viability, are time-consuming and labor-intensive, which limits global genetic studies. Here, we optimized CRISPR interference (CRISPRi...

    Mathilde Guzzo, Lennice K. Castro, Christopher R. Reisch, Monica S. Guo, Michael T. Laub
  • Open Access
    Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and AcrIIC5 Anti-CRISPR Proteins
    Research Article | Molecular Biology and Physiology
    Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and AcrIIC5 Anti-CRISPR Proteins

    As one of their countermeasures against CRISPR-Cas immunity, bacteriophages have evolved natural inhibitors known as anti-CRISPR (Acr) proteins. Despite the existence of such examples for type II CRISPR-Cas systems, we currently know relatively little about the breadth of Cas9 inhibitors, and most of their direct Cas9 targets are uncharacterized. In this work we identify two new type II-C anti-CRISPRs and their cognate Cas9 orthologs,...

    Jooyoung Lee, Aamir Mir, Alireza Edraki, Bianca Garcia, Nadia Amrani, Hannah E. Lou, Ildar Gainetdinov, April Pawluk, Raed Ibraheim, Xin D. Gao, Pengpeng Liu, Alan R. Davidson, Karen L. Maxwell, Erik J. Sontheimer
  • Open Access
    Research Article
    Introduction of Large Sequence Inserts by CRISPR-Cas9 To Create Pathogenicity Mutants in the Multinucleate Filamentous Pathogen Sclerotinia sclerotiorum
    Jingtao Li, Yanhua Zhang, Yucheng Zhang, Pei-Ling Yu, Hongyu Pan, Jeffrey A. Rollins
  • Open Access
    Research Article
    Efficient and Scalable Precision Genome Editing in Staphylococcus aureus through Conditional Recombineering and CRISPR/Cas9-Mediated Counterselection
    Kelsi Penewit, Elizabeth A. Holmes, Kathryn McLean, Mingxin Ren, Adam Waalkes, Stephen J. Salipante
  • Open Access
    Research Article
    Rapid, Selection-Free, High-Efficiency Genome Editing in Protozoan Parasites Using CRISPR-Cas9 Ribonucleoproteins

    Protozoan parasites remain some of the highest-impact human and animal pathogens, with very limited treatment and prevention options. The development of improved therapeutics and vaccines depends on a better understanding of the unique biology of these organisms, and understanding their biology, in turn, requires the ability to track and manipulate the products of genes. In this work, we describe new methods that are available to...

    Lia Carolina Soares Medeiros, Lilith South, Duo Peng, Juan M. Bustamante, Wei Wang, Molly Bunkofske, Natasha Perumal, Fernando Sanchez-Valdez, Rick L. Tarleton
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