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Bacteriophages, or phages, are viruses that specifically infect bacteria, utilizing their genetic material to hijack host cellular machinery for replication. DNA bacteriophages employ single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) genomes. These phages exhibit diverse replication strategies and host interactions, influencing their ecological roles and applications in biotechnology and medicine.ssDNA BacteriophagesssDNA phages, with their small genomes, utilize unique strategies to...
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Bacteriophages, or phages, are viruses that specifically infect bacteria. Among them, T-even bacteriophages, such as T4, exhibit a well-characterized lytic replication cycle in Escherichia coli (E. coli). This process ensures the rapid proliferation of the virus while ultimately leading to the destruction of the bacterial host.Attachment and DNA InjectionThe infection process begins with the recognition and binding of the T4 phage to the E. coli cell surface. Tail fibers of the phage...
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Engineering a Dynamic Controllable Infectivity Switch in Bacteriophage T7.

Chutikarn Chitboonthavisuk1,2,3, Chun Huai Luo1,2, Phil Huss1,2,3

  • 1Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.

ACS Synthetic Biology
|January 5, 2022
PubMed
Summary
This summary is machine-generated.

Synthetic biology enables control of bacteriophage T7 infectivity. Researchers engineered a controllable switch in the phage genome, demonstrating tunable phage activity for potential therapeutic applications.

Keywords:
engineered bacteriophagesenterobacteria phage T7ligand-responsive repression systempseudolysogenic

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Area of Science:

  • Synthetic biology
  • Microbiology
  • Genetics

Background:

  • Transcriptional repressors are key regulators of the bacteriophage T7 life cycle.
  • Engineering bacteriophages offers potential for novel therapeutic and research tools.

Purpose of the Study:

  • To engineer a dynamic and controllable infectivity switch in bacteriophage T7 using synthetic transcription repressors.
  • To demonstrate user control over phage infectivity through a synthetic genetic circuit.

Main Methods:

  • Engineered T7 phage by replacing genomic regions with ligand-responsive promoters and ribosome binding sites (RBS).
  • Controlled phage RNA polymerase (gp1) expression using synthetic repressors.
  • Assessed phage viability, latent period, and titer under repressed and non-repressed conditions.

Main Results:

  • Engineered T7 phages exhibited comparable viability to wildtype when not repressed.
  • An engineered switch utilizing a TetR-responsive promoter and attenuated RBS significantly repressed phage activity.
  • Repression led to a 2-fold increase in latent period and a 10-fold decrease in phage titer.
  • Phage activity was tunable via inducer concentration.

Conclusions:

  • A synthetic genetic circuit can be successfully introduced into the T7 phage genome to create a controllable infectivity switch.
  • This engineered switch allows for user-defined modulation of phage activity.
  • The findings provide a proof of concept for synthetic biology applications in bacteriophage engineering.