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In response to the escalating antibiotic resistance in multidrug-resistant pathogens, we propose an innovative phagemid-based capsid system to generate CRISPR-Cas13a-loaded antibacterial capsids (AB-capsids) for targeted therapy against multidrug-resistant Staphylococcus aureus. Our optimized phagemid system maximizes AB-capsid yield and purity, showing a positive correlation with phagemid copy number. Notably, an 8.65-fold increase in copy number results in a 2.54-fold rise in AB-capsid generation. Phagemids carrying terL-terS-rinA-rinB (prophage-encoded packaging site genes) consistently exhibit high packaging efficiency, and the generation of AB-capsids using lysogenized hosts with terL-terS deletion resulted in comparatively lower level of wild-type phage contamination, with minimal compromise on AB-capsid yield. These generated AB-capsids selectively eliminate S. aureus strains carrying the target gene while sparing non-target strains. In conclusion, our phagemid-based capsid system stands as a promising avenue for developing sequence-specific bactericidal agents, offering a streamlined approach to combat antibiotic-resistant pathogens within the constraints of efficient production and targeted efficacy.
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http://dx.doi.org/10.1038/s42003-024-06754-w | DOI Listing |
Commun Biol
September 2024
Division of Bacteriology, Department of Infection and Immunity, School of Medicine, Jichi Medical University, Shimotsuke city, Tochigi, 329-0498, Japan.
Nucleic Acids Res
June 2003
Crucell Holland B.V., PO Box 2048, 2301CA, Leiden, The Netherlands.
Phage display is a widely used technology for the isolation of peptides and proteins with specific binding properties from large libraries of these molecules. A drawback of the common phagemid/helper phage systems is the high infective background of phages that do not display the protein of interest, but are propagated due to non-specific binding to selection targets. This and the enhanced growth rates of bacteria harboring aberrant phagemids not expressing recombinant proteins leads to a serious decrease in selection efficiency.
View Article and Find Full Text PDFNucleic Acids Res
March 2002
IG Therapy Co., Nong 3rd Building 112, Kangwon National University, Chunchon 200-701, South Korea.
Phage display technology has been applied in many fields of biological and medical sciences to study molecular interactions and especially in the generation of monoclonal antibodies of human origin. However, extremely low display level of antibody molecules on the surface of phage is an intrinsic problem of a phagemid-based display system resulting in low success rate of isolating specific binding molecules. We show here that display of single-chain antibody fragment (scFv) generated with pIGT3 phagemid can be increased dramatically by using a genetically modified Ex-phage.
View Article and Find Full Text PDFChin Med Sci J
March 1998
Beijing Institute for Cancer Research, Beijing 100034.
A library of 2 x 10(7) random octapeptides was constructed by use of phagemid-based monovalent phage display system. The randomly synthesized degenerated oligodeoxyribonucleotides (oligos) were fused to the truncated g III (p230-p403). Sequence analysis of 11 randomly chosen clones suggested that the degenerated inserts and its deduced amino acid (aa) sequences are randomly distributed.
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