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Programming virulent bacteriophages by developing a multiplex genome engineering method.
Hailin Zhang1,2, Ru Zhu1, Zhaofei Wang3
1State Key Laboratory of Microbial Technology, Institute of Microbial Technology, Helmholtz International Lab for Anti-infectives, Shandong University-Helmholtz Institute of Biotechnology, Shandong University, Qingdao, Shandong, China.
Mbio
|May 23, 2025
Summary
We developed a SMART method for engineering virulent bacteriophages (phages). This technique enables multiplex genome modification, creating custom phages to combat bacterial infections effectively.
Area of Science:
- Synthetic biology
- Microbiology
- Genetics
Background:
- Virulent bacteriophages (phages) show promise for combating pathogenic bacteria.
- Engineering virulent phage genomes is challenging due to rapid lysis and toxic gene products.
Purpose of the Study:
- To develop a method for multiplex genome engineering of virulent phages.
- To create a chassis phage and synthetic phages with enhanced lytic capabilities.
Main Methods:
- Developed the SMART (splitting, modifying, assembling, and rebooting) method.
- Engineered the T7 E. coli phage genome by splitting, modifying, assembling, and rebooting.
- Constructed synthetic T7 phages expressing heterologous lysins.
Main Results:
- Successfully deleted 3.9 kb across 8 sites in the T7 phage genome, creating a chassis phage.
- Demonstrated the insertion capacity and expression of exogenous genes in the chassis phage.
- Engineered synthetic T7 phages that efficiently lyse E. coli, S. aureus, and S. agalactiae.
Conclusions:
- The SMART method facilitates multiplex genome engineering of virulent phages.
- Custom-designed phages can be created for therapeutic applications with enhanced efficacy and specificity.
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