Optimized programmable control of biomaterial-dependent bacterial adhesion for a clinical uropathogen using CRISPR interference

ABSTRACT: Biofilm formation on indwelling medical devices and biomaterial surfaces poses a major threat to human health by driving persistent and treatment-resistant infections. However, the genetic mechanisms through which surface chemistry shapes the determinants of early-stage bacterial adhesion on biomaterials remain poorly resolved. Here, using a hydrophobic silicone (polydimethylsiloxane, PDMS) and a hydrophilic hydrogel (polyethylene glycol dimethacrylate, PEG) as contrasting biomaterial models for urinary catheter coatings, we employed CRISPR interference (CRISPRi) to study the effects of targeted gene repression on early-stage attachment of uropathogenic Escherichia coli CFT073. Screening identified multiple genes that elicit material-dependent adhesion phenotypes upon CRISPRi silencing. While repression of many canonical adhesion determinants consistently reduced attachment across both substrates, repression of lpcA produced diametric adhesion to PDMS and PEG. Multiplexed CRISPRi targeting of up to six canonical adhesion determinants revealed material-dependent patterns of adhesion suppression. A genome-wide CRISPRi dataset was applied to identify gene targets that are non-conical adhesion determinants, including those that confer material-dependent control of adhesion. CRISPRi targeting of an uncharacterized protein (yejL) yielded the greatest adhesion suppression, and RNA-seq profiling revealed extensive transcriptome remodeling spanning 2,284 and 1,813 differentially expressed genes on PDMS and PEG, respectively. Collectively, these findings elucidate gene targets to control uropathogen adhesion, provide a material-synthetic biology framework to study bacterial adhesion, and support the biomaterial context-dependent nature of bacterial adhesion that emerges from the interplay between surface physicochemistry and gene regulation.

ORGANISM(S): Escherichia coli

PROVIDER: GSE349377 | GEO | 2026/10/22

REPOSITORIES: GEO