RNA toehold switch-based reporter assay to assess bacterial uptake of antisense oligomers.

ABSTRACT: Antisense oligomers (ASOs) hold promise as antibiotics for the selective targeting of bacterial pathogens and as tools for the modulation of gene expression in microbes that are not amenable to genetic engineering. However, their efficient delivery across the complex bacterial envelope remains a major challenge. There are few methods to assess the efficiency of carrier-mediated ASO uptake by bacteria. Here, we have developed a "switch-on" reporter assay to measure ASO uptake efficiency in a semi-quantitative manner. The assay uses a synthetic RNA toehold switch fused to the mRNA of a fluorescent reporter protein, which is activated in vivo by a peptide nucleic acid (PNA)-based ASO upon delivery into the bacterial cytosol. We have used this assay to screen different cell-penetrating

SUBMITTER: Sarkar P 

PROVIDER: S-EPMC12017328 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature