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CRISPR

  • Portable CRISPR-Cas9<sup>N</sup> System for Flexible Genome Engineering in <span class="named-content genus-species" id="named-content-1">Lactobacillus acidophilus</span>, <span class="named-content genus-species" id="named-content-2">Lactobacillus gasseri</span>, and <span class="named-content genus-species" id="named-content-3">Lactobacillus paracasei</span>
    Methods
    Portable CRISPR-Cas9N System for Flexible Genome Engineering in Lactobacillus acidophilus, Lactobacillus gasseri, and Lactobacillus paracasei

    This work describes the development of a lactobacillus CRISPR-based editing system for genome manipulations in three Lactobacillus species belonging to the lactic acid bacteria (LAB), which are commonly known for their long history of use in food fermentations and as indigenous members of healthy microbiotas and for their emerging roles in human and animal commercial health-promoting applications. We exploited the established...

    Yong Jun Goh, Rodolphe Barrangou
  • Open Access
    Harnessing CRISPR-Cas9 for Genome Editing in <span class="named-content genus-species" id="named-content-1">Streptococcus pneumoniae</span> D39V
    Methods
    Harnessing CRISPR-Cas9 for Genome Editing in Streptococcus pneumoniae D39V

    Streptococcus pneumoniae (the pneumococcus) is an important opportunistic human pathogen killing more than 1 million people each year. Having the availability of a system capable of easy genome editing would significantly facilitate drug discovery and efforts to identify new vaccine candidates.

    Dimitra Synefiaridou, Jan-Willem Veening
  • Open Access
    A CRISPR Interference Platform for Selective Downregulation of Gene Expression in <span class="named-content genus-species" id="named-content-1">Borrelia burgdorferi</span>
    Methods | Spotlight
    A CRISPR Interference Platform for Selective Downregulation of Gene Expression in Borrelia burgdorferi

    Gene function studies are facilitated by the availability of rapid and easy-to-use genetic tools. Homologous recombination-based methods traditionally used to genetically investigate gene function remain cumbersome to perform in B. burgdorferi, as they often are relatively inefficient.

    Constantin N. Takacs, Molly Scott, Yunjie Chang, Zachary A. Kloos, Irnov Irnov, Patricia A. Rosa, Jun Liu, Christine Jacobs-Wagner
  • Open Access
    Fragment Exchange Plasmid Tools for CRISPR/Cas9-Mediated Gene Integration and Protease Production in <span class="named-content genus-species" id="named-content-1">Bacillus subtilis</span>
    Biotechnology
    Fragment Exchange Plasmid Tools for CRISPR/Cas9-Mediated Gene Integration and Protease Production in Bacillus subtilis

    We complemented a cloning platform with new editing plasmids that allow a quick transition from high-throughput cloning and the expression of new enzymes to the stable integration of genes for the production of enzymes through B. subtilis fermentation. We present two systems for the effective assembly cloning of any genome-editing cassette that shortens the...

    Antonio García-Moyano, Øivind Larsen, Sushil Gaykawad, Eleni Christakou, Catherine Boccadoro, Pål Puntervoll, Gro Elin Kjæreng Bjerga
  • Novel Genus of Phages Infecting <span class="named-content genus-species" id="named-content-1">Streptococcus thermophilus</span>: Genomic and Morphological Characterization
    Food Microbiology
    Novel Genus of Phages Infecting Streptococcus thermophilus: Genomic and Morphological Characterization

    Despite decades of research and adapted antiphage strategies such as CRISPR-Cas systems, virulent phages are still a persistent risk for the milk fermentation industry worldwide, as they can cause manufacturing failures and alter product quality. Phages P738 and D4446 are novel virulent phages that infect the food-grade Gram-positive bacterial species Streptococcus...

    Cécile Philippe, Sébastien Levesque, Moïra B. Dion, Denise M. Tremblay, Philippe Horvath, Natascha Lüth, Christian Cambillau, Charles Franz, Horst Neve, Christophe Fremaux, Knut J. Heller, Sylvain Moineau
  • Using an Endogenous CRISPR-Cas System for Genome Editing in the Human Pathogen <span class="named-content genus-species" id="named-content-1">Clostridium difficile</span>
    Genetics and Molecular Biology | Spotlight
    Using an Endogenous CRISPR-Cas System for Genome Editing in the Human Pathogen Clostridium difficile

    Clostridium difficile represents today a real danger for human and animal health. It is the leading cause of diarrhea associated with health care in adults in industrialized countries. The incidence of these infections continues to increase, and this trend is accentuated by the general aging of the population. Many questions about the mechanisms contributing to...

    Anna Maikova, Victor Kreis, Anaïs Boutserin, Konstantin Severinov, Olga Soutourina
  • Open Access
    Determinants of Phage Host Range in <em>Staphylococcus</em> Species
    Minireview | Spotlight
    Determinants of Phage Host Range in Staphylococcus Species

    Bacteria in the genus Staphylococcus are important targets for phage therapy due to their prevalence as pathogens and increasing antibiotic resistance. Here we review Staphylococcus outer surface features and specific phage resistance mechanisms that define the host range, the set of strains that an individual phage can potentially infect.

    Abraham G. Moller, Jodi A. Lindsay, Timothy D. Read
  • CRISPR-Cas9 and CRISPR-Assisted Cytidine Deaminase Enable Precise and Efficient Genome Editing in <span class="named-content genus-species" id="named-content-1">Klebsiella pneumoniae</span>
    Methods
    CRISPR-Cas9 and CRISPR-Assisted Cytidine Deaminase Enable Precise and Efficient Genome Editing in Klebsiella pneumoniae

    Genetics is a key means to study bacterial physiology. However, the highly desirable scarless genetic manipulation is often time-consuming and laborious for the major human pathogen K. pneumoniae. We developed a CRISPR-Cas9-mediated genome-editing method and a cytidine base-editing system, enabling rapid, highly efficient, and iterative genome editing in both...

    Yu Wang, Shanshan Wang, Weizhong Chen, Liqiang Song, Yifei Zhang, Zhen Shen, Fangyou Yu, Min Li, Quanjiang Ji
  • High-Resolution Identification of Multiple <span class="named-content genus-species" id="named-content-1">Salmonella</span> Serovars in a Single Sample by Using CRISPR-SeroSeq
    Food Microbiology | Spotlight
    High-Resolution Identification of Multiple Salmonella Serovars in a Single Sample by Using CRISPR-SeroSeq

    Salmonella enterica is the leading bacterial cause of foodborne illness in the United States and is represented by over 2,600 distinct serovars. Some of these serovars are pathogenic in humans, while others are not. Current surveillance for this pathogen is limited by the detection of only the most abundant serovars, due to the culture-based approaches that are used....

    Cameron P. Thompson, Alexandra N. Doak, Naufa Amirani, Erin A. Schroeder, Justin Wright, Subhashinie Kariyawasam, Regina Lamendella, Nikki W. Shariat
  • Methods
    CRISPR-Cpf1-Assisted Multiplex Genome Editing and Transcriptional Repression in Streptomyces

    Rapid, efficient genetic engineering of Streptomyces strains is critical for genome mining of novel natural products (NPs) as well as strain improvement. Here, a novel and high-efficiency Streptomyces genome editing tool is established based on the FnCRISPR-Cpf1 system, which is an...

    Lei Li, Keke Wei, Guosong Zheng, Xiaocao Liu, Shaoxin Chen, Weihong Jiang, Yinhua Lu

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