Speaker
Description
Cell-penetrating peptides (CPPs) can be harnessed to deliver antisense oligomers (ASOs) into bacterial cells. However, CPPs that are efficiently taken up by Bacteroides—an abundant species of the human gut microbiota—are not currently known. Here, we perform a global screen for CPP uptake by Bacteroides thetaiotaomicron. We perform a global screen for bacterial cell-penetrating peptides (CPPs), identifying MK2i (mitogen-activated protein kinase-activated protein kinase 2 inhibitory peptide) as a suitable carrier peptide with high uptake efficiency and specificity. Our data further propose a TonB-dependent import mechanism and reveal influences of media composition, growth phase, and cell surface structure on CPP uptake efficiency. Building on these results, we design an MK2i-coupled to an ASO peptide nucleic acid (PNA) against fabG, an essential Bacteroides gene involved in envelope fatty acid synthesis. Low micromolar concentrations of this CPP-PNA conjugate evoke a dose-dependent growth inhibition and bactericidal effects. Leveraging a fluorescent B. thetaiotaomicron reporter strain, we monitor the effect of PNA-targeting on GFP transcript, protein, and fluorescence levels. These findings lay the groundwork for delivering ASOs into and leveraging ASO technology in a microbiota keystone species.
Keywords
antisense oligomer; cell-penetrating peptide; peptide nucleic acid; capsular polysaccharide; MK2i; FabG; Bacteroidota
| Registration ID | #177 |
|---|---|
| Professional Status of the Speaker | PhD Student |
| Junior Scientist Status | Yes, I am a Junior Scientist. |
Author
Co-authors
External references
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