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Mitigating the attachment of microorganisms to polymer biomaterials is critical for preventing hospital-acquired infections. Two chemical strategies to mitigate fouling include fabricating fouling-resistant surfaces, which typically present hydrophilic polymers, such as polyethylene glycol (PEG), or creating fouling-release surfaces, which are generally hydrophobic featuring polydimethylsiloxane (PDMS). Despite the demonstrated promise of employing PEG or PDMS, amphiphilic PEG/PDMS copolymer materials remain understudied. Here, for the first time, we investigated if phase-separated amphiphilic copolymers confounded microbial adhesion. We used bottlebrush amphiphilic PEG/PDMS co-networks and homopolymer networks to study bacterial adhesion across a library of gels (ϕPEG = 0.00, 0.21, 0.40, 0.55, 0.80, and 1.00). Hydrated atomic force microscopy measurements revealed that most of the gels had low surface roughness, less than 5 nm, and an elastic modulus of ∼80 kPa. Interestingly, the surface roughness and elastic modulus of the ϕPEG = 0.40 gel were twice as high as those of the other gels due to the presence of crystalline domains, as confirmed using polarized optical microscopy on the hydrated gel. The interactions of these six well-characterized gels with bacteria were determined using Escherichia coli K12 MG1655 and Staphylococcus aureus SH1000. The attachment of both microbes decreased by at least 60% on all polymer gels versus the glass controls. S. aureus adhesion peaked on the ϕPEG = 0.40, likely due to its increased elastic modulus, consistent with previous literature demonstrating that modulus impacts microbial adhesion. These findings suggest that hydrophilic, hydrophobic, and amphiphilic biomaterials effectively resist the early attachment of Gram-negative and Gram-positive microorganisms, providing guidance for the design of next-generation antifouling surfaces. 

CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) have become ubiquitous approaches to control gene expression in bacteria due to their simple design and effectiveness. By regulating transcription of a target gene(s), CRISPRi/a can dynamically engineer cellular metabolism, implement transcriptional regulation circuitry, or elucidate genotype-phenotype relationships from smaller targeted libraries up to whole genome-wide libraries. While CRISPRi/a has been primarily established in the model bacteria Escherichia coli and Bacillus subtilis , a growing numbering of studies have demonstrated the extension of these tools to other species of bacteria (here broadly referred to as non-model bacteria). In this mini-review, we discuss the challenges that contribute to the slower creation of CRISPRi/a tools in diverse, non-model bacteria and summarize the current state of these approaches across bacterial phyla. We find that despite the potential difficulties in establishing novel CRISPRi/a in non-model microbes, over 190 recent examples across eight bacterial phyla have been reported in the literature. Most studies have focused on tool development or used these CRISPRi/a approaches to interrogate gene function, with fewer examples applying CRISPRi/a gene regulation for metabolic engineering or high-throughput screens and selections. To date, most CRISPRi/a reports have been developed for common strains of non-model bacterial species, suggesting barriers remain to establish these genetic tools in undomesticated bacteria. More efficient and generalizable methods will help realize the immense potential of programmable CRISPR-based transcriptional control in diverse bacteria.