An improved approach was developed for the genetic manipulation of the Gram-positive extremophile (formerly called ). We describe an allelic replacement method originally developed for that allows the deletion, mutation, or insertion of genes without leaving markers or other genetic scars. In addition, a protocol for rapid plasmid methylation and transformation is presented. The combined methods allow the routine genetic manipulation of from initial transformation to the desired strain in 8 days. These methods improve as a model organism for the study of extremophiles.
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http://dx.doi.org/10.3389/fmicb.2024.1465811 | DOI Listing |
mBio
January 2025
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, Texas, USA.
The chick embryo chorioallantoic membrane (CAM) tumor model is a valuable preclinical model for studying the tumor-colonizing process of serovar Typhimurium. It offers advantages such as cost-effectiveness, rapid turnaround, reduced engraftment issues, and ease of observation. In this study, we explored and validated the applicability of the partially immune-deficient CAM tumor model.
View Article and Find Full Text PDFLife Metab
February 2025
New Cornerstone Science Laboratory, State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, National Biomedical Imaging Center, The Beijing Laboratory of Biomedical Imaging, Peking-Tsinghua Center for Life Sciences, School of Future Technology, Peking University, Beijing 100871, China.
Glucose-stimulated insulin release from pancreatic β-cells is critical for maintaining blood glucose homeostasis. An abrupt increase in blood glucose concentration evokes a rapid and transient rise in insulin secretion followed by a prolonged, slower phase. A diminished first phase is one of the earliest indicators of β-cell dysfunction in individuals predisposed to develop type 2 diabetes.
View Article and Find Full Text PDFBio Protoc
January 2025
Department of Cell Biology and Human Anatomy, University of California, Davis, School of Medicine, Davis, CA, USA.
Mouse embryonic fibroblasts (MEFs) derived from genetically modified mice are a valuable resource for studying gene function and regulation. The MEF system can also be combined with rescue studies to characterize the function of mutant genes/proteins, such as disease-causing variants. However, primary MEFs undergo senescence soon after isolation and passaging, making long-term genetic manipulations difficult.
View Article and Find Full Text PDFBiotechnol Bioeng
January 2025
Department of Biosystems Engineering, Auburn University, Auburn, Alabama, USA.
Bacillus velezensis FZB42 is a prominent plant growth-promoting rhizobacterium and biocontrol agent known for producing a wide array of antimicrobial compounds. The capability to genetically manipulate this strain would facilitate understanding its metabolism and enhancing its sustainable agriculture applications. In this study, we report the first successful implementation of high-efficiency CRISPR-Cas9 genome editing in B.
View Article and Find Full Text PDFNat Commun
January 2025
Institute for Advanced Biosciences, Team: Epigenetics, Immunity, Metabolism, Cell Signaling & Cancer, Inserm U 1209, CNRS UMR 5309, Univ. Grenoble Alpes, Grenoble, France.
Dendritic cells (DC) are key players in antitumor immune responses. Tumors exploit their plasticity to escape immune control; their aberrant surface carbohydrate patterns (e.g.
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