Abstract
BackgroundStaphylococcus aureus (S. aureus) has the potential to opportunistically cause infectious diseases, including osteomyelitis, skin infections, pneumonia, and diarrhea. We previously reported that ASyycG RNA reduced the transcripts of virulent genes, and biofilm formation of S. aureus. Currently, graphene oxide (GO) nanosheets are used to efficiently deliver nucleic acids with favorable biocompatibility.MethodsIn the current study, a GO-based recombinant pDL278 ASyycG vector transformation strategy was developed. The particle size distributions and zeta-potential of the GO-PEI-based ASyycG were evaluated. The ASyycG plasmids were labeled with gene-encoding enhanced green fluorescent protein (ASyycG-eGFP). Quantitative real-time PCR assays were performed to investigate the expression of yycF/G/H and icaADB genes. Biofilm biomass and bacterial viability of S. aureus were evaluated by scanning electron microscopy and confocal laser scanning microscopy. We found that the expression of the yycG gene was inversely correlated with levels of the ASyycG transcripts and that the GO-PEI-ASyycG strain had the lowest expression of biofilm organization-associated genes.ResultsThe results showed that the GO-based strategy significantly increased ASyycG transformation as a delivery system compared to the conventional competence-stimulating peptide strategy. Furthermore, GO-PEI-ASyycG suppressed bacterial biofilm aggregation and improved bactericidal effects on S. aureus after 24 h biofilm establishment.ConclusionsOur findings demonstrated that nano-GO with antisense yycG RNA is a more effective and relatively stable strategy for the management of S. aureus infections.
Highlights
Staphylococcus aureus (S. aureus), a gram-positive coccus, is carried by about 20–30% of healthy individuals and mostly colonizes the nasopharynx [1]
graphene oxide (GO)-PEI-antisense yycG sequences (ASyycG) increased ASyycG transformation and significantly reduced biofilm formation-associated gene expression Using confocal laser scanning microscopy, higher levels of GFP-expression were observed in samples induced with GO-PEI-ASyycG compared to pure ASyycG (Fig. 2a)
Quantitative Real-time polymerase chain reaction (RT-PCR) assays demonstrated that expression levels of ASyycG RNA in the ASyycG and GO-PEI-ASyycG strains were significantly increased 2.8-fold and 6.5-fold, respectively, compared to S. aureus ATCC29213 strains (n = 10, p < 0.05; Fig. 2c)
Summary
Staphylococcus aureus (S. aureus), a gram-positive coccus, is carried by about 20–30% of healthy individuals and mostly colonizes the nasopharynx [1]. S. aureus has the potential to cause a wide range of diseases, including osteomyelitis, skin infections, pneumonia, and even life-threatening infective endocarditis associated with considerable global human morbidity and mortality [2]. In some cases, S. aureus is resistant to multiple types of antibiotics, which has been attributed to the abuse of antibiotics, resulting in the emergence of methicillin-resistant Staphylococcus aureus (MRSA) [3]. The sharp edges on the GO sheet structures physically disrupt cell membranes and cause oxidative stress reactions. GO sheets can be used to effectively deliver single-stranded nucleic acids, the ability of GO to carry double-stranded DNA (dsDNA) is limited. Staphylococcus aureus (S. aureus) has the potential to opportunistically cause infectious diseases, including osteomyelitis, skin infections, pneumonia, and diarrhea. Graphene oxide (GO) nanosheets are used to efficiently deliver nucleic acids with favorable biocompatibility
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