Mycoplasmas are minimal bacteria that infect humans, wildlife, and most economically relevant livestock species. Mycoplasma infections cause a large range of chronic inflammatory diseases, eventually leading to death in some animals. Due to the lack of efficient recombination and genome engineering tools for most species, the production of mutant strains for the identification of virulence factors and the development of improved vaccine strains is limited. Here, we demonstrate the adaptation of an efficient Cas9-Base Editor system to introduce targeted mutations into three major pathogenic species that span the phylogenetic diversity of these bacteria: the avian pathogen Mycoplasma gallisepticum and the two most important bovine mycoplasmas, Mycoplasma bovis and Mycoplasma mycoides subsp. . As a proof of concept, we successfully used an inducible SpdCas9-pmcDA1 cytosine deaminase system to disrupt several major virulence factors in these pathogens. Various induction times and inducer concentrations were evaluated to optimize editing efficiency. The optimized system was powerful enough to disrupt 54 of 55 insertion sequence transposases in a single experiment. Whole-genome sequencing of the edited strains showed that off-target mutations were limited, suggesting that most variations detected in the edited genomes are Cas9-independent. This effective, rapid, and easy-to-use genetic tool opens a new avenue for the study of these important animal pathogens and likely the entire class . Mycoplasmas are minimal pathogenic bacteria that infect a wide range of hosts, including humans, livestock, and wild animals. Major pathogenic species cause acute to chronic infections involving still poorly characterized virulence factors. The lack of precise genome editing tools has hampered functional studies of many species, leaving multiple questions about the molecular basis of their pathogenicity unanswered. Here, we demonstrate the adaptation of a CRISPR-derived base editor for three major pathogenic species: Mycoplasma gallisepticum, Mycoplasma bovis, and Mycoplasma mycoides subsp. Several virulence factors were successfully targeted, and we were able to edit up to 54 target sites in a single step. The availability of this efficient and easy-to-use genetic tool will greatly facilitate functional studies of these economically important bacteria.
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http://dx.doi.org/10.1128/aem.00996-22 | DOI Listing |
China CDC Wkly
January 2025
Department of Clinical Laboratory, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou City, Zhejiang Province, China.
Introduction: Type F () represents a significant pathogen in human gastrointestinal diseases, primarily through its gene encoding enterotoxin (CPE). This investigation examined the prevalence, antimicrobial resistance patterns, and genetic characteristics of Type F within the Chinese population.
Methods: The study analyzed 2,068 stool samples collected from 11 provincial hospitals in 2024.
Cell Surf
June 2025
Departamento de Biología, División de Ciencias Naturales y Exactas, Campus Guanajuato, Universidad de Guanajuato, Noria Alta s/n, col. Noria Alta, C.P. 36050 Guanajuato, Gto, Mexico.
is one of the leading etiological agents of sporotrichosis, a cutaneous and subcutaneous mycosis worldwide distributed. This organism has been recently associated with epidemic outbreaks in Brazil. Despite the medical relevance of this species, little is known about its virulence factors, and most of the information on this subject is extrapolated from .
View Article and Find Full Text PDFJ Vet Diagn Invest
January 2025
Asia-Pacific Centre for Animal Health, Melbourne Veterinary School, Faculty of Science, University of Melbourne, Werribee, Victoria, Australia.
The complex contains important opportunistic pathogens of humans and vertebrate animals, as well as insects and other invertebrates. To date, the methods used for the identification of species within the genus , including PCR assays, have poor discriminatory power and may require further molecular typing or genomic sequence analysis to determine clinical relevance. We developed a duplex TaqMan probe-based quantitative real-time PCR (qPCR) assay targeting the gene, which is involved in chitin degradation and transport, and the gene, which is involved in urease production.
View Article and Find Full Text PDFBiochemistry (Mosc)
December 2024
Faculty of Biology, St. Petersburg State University, St. Petersburg, 199034, Russia. ARRAY(0x5ae2b7af6df8).
Amyloids are protein fibrils with a characteristic cross-β structure that is responsible for the unusual resistance of amyloids to various physical and chemical factors, as well as numerous pathogenic and functional consequences of amyloidogenesis. The greatest diversity of functional amyloids was identified in bacteria. The majority of bacterial amyloids are involved in virulence and pathogenesis either via facilitating formation of biofilms and adaptation of bacteria to colonization of a host organism or through direct regulation of toxicity.
View Article and Find Full Text PDFBioorg Chem
January 2025
Helmholtz International Lab for Anti-Infectives, State Key Laboratory of Microbial Technology, Shandong University, Qingdao, Shandong 266237, China. Electronic address:
Infections of multidrug-resistant pathogens including Pseudomonas aeruginosa, cause a high risk of mortality in immunocompromised patients and underscore the need for novel natural antibacterial drugs. In this study, common phytochemicals prevalent in fruits and vegetables have been demonstrated for their ability to inhibit quorum sensing (QS) in Pseudomonas aeruginosa PAO1 (PA). Ten compounds were screened virtually by molecular docking, among which, daidzein dimethyl ether originally from Albizzia lebbeck showed the most significant inhibitory effect on the formation of biofilm and the accumulation of virulence factors, including elastase, pyocyanin and rhamnolipid in PA.
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