{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T01:42:05Z","timestamp":1768527725919,"version":"3.49.0"},"posted":{"date-parts":[[2025,10,15]]},"group-title":"Pharmacology and Toxicology","reference-count":39,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T00:00:00Z","timestamp":1760486400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institute of Allergy and Infectious Diseases of the National Institutes of Health","award":["R16AI184350"],"award-info":[{"award-number":["R16AI184350"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,10,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Bacteriophages offer advantages over small-molecule antibiotics, including host specificity and general compatibility with the human phagenome. However, their evolvability as replicating biological entities introduces therapeutic unpredictability and risks of phage-bacteria co-evolution. Here, we retain the targeting benefits of phages while avoiding genetic replication by engineering genetic-information-free, protein-only phages (POPs). These genome-free particles self-assemble in a cell-free protein synthesis system from modular, de novo gene fragments encoding only structural and antimicrobial proteins. Using\n                  <jats:italic>Enterobacteria phage<\/jats:italic>\n                  T7 and its susceptible bacterial host as a model, we test the hypothesis that POPs stochastically encapsulate small antimicrobial proteins during self-assembly and deliver them into bacteria during adsorption via an ejectome-mediated injection mechanism. A computational survey of T7 small proteins revealed early and mid-genome enrichments of hypothetical proteins and capsid volume sufficient to accommodate multiple small proteins in the absence of the \u223c40-kb genome. In time-series antimicrobial susceptibility assays (48-72 h), POPs produced initial growth inhibition comparable to wild-type T7 at the highest doses with a linear dose-effect relationship and a minimum inhibitory concentration-like threshold. These results establish the feasibility of genetic-information-free POPs as protein-based antimicrobials that couple phage receptor specificity with minimal biosafety risks, supporting the development of more stable and predictable phage-inspired therapeutics.\n                <\/jats:p>","DOI":"10.1101\/2025.10.13.682247","type":"posted-content","created":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T14:20:11Z","timestamp":1760538011000},"source":"Crossref","is-referenced-by-count":0,"title":["Targeting multidrug-resistant bacteria with genetic-information-free protein-only phages"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7663-4611","authenticated-orcid":false,"given":"Jeanette","family":"Batres","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-4490-8307","authenticated-orcid":false,"given":"Branden","family":"Hunter","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7052-0971","authenticated-orcid":false,"given":"Hyunjin","family":"Shim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"issue":"10325","key":"2025101723451099000_2025.10.13.682247v1.1","first-page":"629","article-title":"Global burden of bacterial antimicrobial resistance in 2019","volume":"399","year":"2022","journal-title":"a systematic analysis. 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