Finding novel, innovative and sustainable ways to preserve the efficacy of currently-licensed antimicrobial drugs is a central part of efforts to address the antimicrobial resistance (AMR) crisis. Anti-sense oligonucleotides (ASOs) are short oligonucleotides that can provide a pathway for gene silencing by RNase H degradation. Recent improvements in the design and chemistry of ASOs have opened up a new outlet for this technology as tools for basic research and drug discovery. The overarching objective of this project is to evaluate the hypothesis that ASOs have therapeutic potential as inhibitors of growth and resistance in ESKAPE pathogens. In order to test this, preliminary bioinformatic analysis, disk diffusions assays and MICs were carried out to characterise the resistance profiles of the ESKAPE pathogens used in this study. To deliver ASOs into cells, cell-penetrating oligopeptides (CPPs) were synthesised using Fmoc solid-phase peptide synthesis with specific linkers. Confocal microscopy and flow cytometry is being used to evaluate the incorporation rate and percentage of fluorescently labelled CPPs into cells of Gram-negative and Gram-positive ESKAPE pathogens. pVec (a CPP) has been synthesised with different linkers to date. Conjugation experiments are underway to establish optimal click-chemistry conditions for biological applications. Susceptibility tests carried out to date revealed some highly resistant strains, and the genes responsible for these resistances are being evaluated further, through bioinformatics analyses, as potential ASO targets. This multidisciplinary project aims to enable the re-purposing of antibiotics as part of efforts to overcome resistance in the ESKAPE pathogens.