Identification of Antibacterial Cyclic Peptides with a High-Throughput Cell-Based Dropout Screen
Abstract
Antimicrobial resistance is a growing global health threat, necessitating new antibiotics and discovery strategies. Here, we report a target-agnostic negative-selection screening platform that combines an intracellular library of 3.2 million cyclic peptides with next-generation sequencing to identify antibacterial cyclic peptides through the depletion of their encoding sequences. Clustering of depleted sequences by shared pharmacophores enables robust hit identification and allows focused library design. Using this approach, we identify a cyclic peptide scaffold that inhibits the previously untargeted essential iron-sulfur cluster carrier protein ErpA. This work establishes a general and scalable framework for the discovery of intracellular antibacterial targets and associated inhibitors directly in living cells.
Fields of Study & Subjects
Chemical Synthesis and Analysis
Antimicrobial Peptides and Activities
Microbial Natural Products and Biosynthesis
Publication details
- Journal / source
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Volume
- 148
- Issue
- 30
- Pages
- 32451–32460
- Publication date
- 25 July 2026
- Language
- EN