TY - JOUR T1 - Engineering Phage Host-Range and Suppressing Bacterial Resistance Through Phage Tail Fiber Mutagenesis JF - bioRxiv DO - 10.1101/699090 SP - 699090 AU - Kevin Yehl AU - Sébastien Lemire AU - Andrew C. Yang AU - Hiroki Ando AU - Mark Mimee AU - Marcelo Der Torossian Torres AU - Cesar de la Fuente-Nunez AU - Timothy K. Lu Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/07/11/699090.abstract N2 - The rapid emergence of antibiotic-resistant infections is prompting increased interest in phage-based antimicrobials. However, acquisition of resistance by bacteria is a major issue in the successful development of phage therapies. Through natural evolution and structural modeling, we identified host-range determining regions (HRDR) in the T3 phage tail fiber protein and developed a high-throughput strategy to genetically engineer these regions through site-directed mutagenesis. Inspired by antibody specificity engineering, this approach generates deep functional diversity (>107 different members), while minimizing disruptions to the overall protein structure, resulting in synthetic “phagebodies”. We showed that mutating HRDRs yields phagebodies with altered host-ranges. Select phagebodies enable long-term suppression of bacterial growth by preventing the appearance of resistance in vitro and are functional in vivo using a mouse skin infection model. We anticipate this approach may facilitate the creation of next-generation antimicrobials that slow resistance development and could be extended to other viral scaffolds for a broad range of applications.HighlightsVastly diverse phagebody libraries containing 107 different members were created.Structure-informed engineering of viral tail fibers efficiently generated host-range alterations.Phagebodies prevented the development of bacterial resistance across long timescales in vitro and are functional in vivo. ER -