Reprogramming Bacteriophage Host Range through Structure-Guided Design of Chimeric Receptor Binding Proteins.

Area of Science:

• Microbiology
• Synthetic Biology
• Structural Biology

Background:

• Bacteriophages are valuable for diagnostics and microbiome manipulation.
• Isolating phages with specific host ranges is a significant challenge.

Purpose of the Study:

• To engineer bacteriophage host range through modification of receptor binding proteins (RBPs).
• To develop a synthetic biology framework for creating phages with tunable host specificities.

Main Methods:

• Identified Listeria phage PSA's RBP (Gp15) and created a randomized RBP library.
• Isolated host range mutants and integrated them into a synthetic polyvalent phage.
• Determined the crystal structure of the Gp15 RBP carboxyl terminus.
• Used bioinformatics and structure-guided design to create chimeric RBPs.

Main Results:

• Successfully engineered a synthetic phage with an extended host range.
• Demonstrated predictable host range expansion through RBP domain exchange.
• Generated chimeric RBPs with modified head, neck, or shoulder domains.

Conclusions:

• Developed a synthetic biology blueprint for bacteriophage host range engineering.
• Structure-guided design facilitates the creation of phages with tailored host specificities.
• This approach enables the development of phage biologics with customizable properties.