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Related Concept Videos

DNA Bacteriophages01:26

DNA Bacteriophages

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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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Lytic Cycle of Bacteriophages01:30

Lytic Cycle of Bacteriophages

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Bacteriophages, also known as phages, are specialized viruses that infect bacteria. A key characteristic of phages is their distinctive “head-tail” morphology. A phage begins the infection process (i.e., lytic cycle) by attaching to the outside of a bacterial cell. Attachment is accomplished via proteins in the phage tail that bind to specific receptor proteins on the outer surface of the bacterium. The tail injects the phage’s DNA genome into the bacterial cytoplasm. In the...
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Viral Replication: Lytic Cycle01:20

Viral Replication: Lytic Cycle

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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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Lysogenic Cycle of Bacteriophages00:43

Lysogenic Cycle of Bacteriophages

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In contrast to the lytic cycle, phages infecting bacteria via the lysogenic cycle do not immediately kill their host cell. Instead, they combine their genome with the host genome, allowing the bacteria to replicate the phage DNA along with the bacterial genome. The incorporated copy of the phage genome is called the prophage. Some prophages can re-activate and enter the lytic cycle. This often occurs in response to a perturbation, such as DNA damage, but can also transpire in the absence of...
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Viral Replication: Lysogenic Cycle01:16

Viral Replication: Lysogenic Cycle

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The lysogenic cycle is a crucial viral replication strategy that allows bacteriophages to persist within host cells without immediately destroying them. This process is primarily observed in temperate phages, such as bacteriophage lambda (λ), which infects Escherichia coli. The cycle allows the viral genome to persist across bacterial generations while keeping host cells viable.Integration of the Viral GenomeUpon infection, bacteriophage lambda attaches to the bacterial surface and injects...
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CRISPR and crRNAs02:53

CRISPR and crRNAs

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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High-throughput approaches to understand and engineer bacteriophages.

Phil Huss1, Jackie Chen2, Srivatsan Raman3

  • 1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA; Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, WI 53706, USA.

Trends in Biochemical Sciences
|September 30, 2022
PubMed
Summary

New high-throughput genome engineering methods enable rapid characterization of bacteriophages (phages). These tools accelerate understanding of how phage sequence dictates function, advancing biological research.

Keywords:
deep sequencingengineered bacteriophagesgenome engineeringphage libraryphage mutagenesisphage–host interactions

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Area of Science:

  • Microbiology
  • Genomics
  • Molecular Biology

Background:

  • Bacteriophage (phage) research is crucial for fundamental biology.
  • Metagenomics has uncovered numerous uncharacterized phages, but understanding their function remains a challenge.
  • Current methods for phage characterization are low-throughput and struggle to link sequence to function.

Purpose of the Study:

  • To introduce powerful emerging technologies for high-throughput genome engineering in phages.
  • To enable rapid characterization of sequence-function relationships in bacteriophages.
  • To outline basic science and engineering applications of these novel approaches.

Main Methods:

  • High-throughput genome engineering techniques.
  • Pooled selection experiments.
  • Deep sequencing for variant analysis.

Main Results:

  • Ability to characterize up to 10^5 phage variants simultaneously.
  • Facilitation of rapid analysis of sequence-function relationships.
  • Demonstration of feasibility for basic science and engineering goals.

Conclusions:

  • Emerging high-throughput genome engineering technologies significantly advance bacteriophage research.
  • These methods overcome limitations of traditional approaches for characterizing phage function.
  • The described techniques open new avenues for exploring phage biology and applications.