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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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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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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
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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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Synthesis of Infectious Bacteriophages in an E. coli-based Cell-free Expression System
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Approaches for bacteriophage genome engineering.

Marina Mahler1, Ana Rita Costa2, Sam P B van Beljouw2

  • 1Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand; Department of Bionanoscience, Delft University of Technology, Delft, the Netherlands.

Trends in Biotechnology
|September 18, 2022
PubMed
Summary

Bacteriophage genome engineering is advancing with new cell-free methods and CRISPR-Cas counter-selection. These techniques overcome challenges in manipulating phage DNA for applications like phage therapy and biocontrol.

Keywords:
CRISPR-Cashomologous recombinationphage engineeringrebootingrecombineering

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Phage therapy shows promise for antibiotic-resistant infections.
  • Phages and their proteins have applications in biocontrol and diagnostics.
  • Phage genome manipulation is hindered by a lack of universal selectable markers.

Purpose of the Study:

  • To review state-of-the-art techniques for engineering bacteriophage genomes.
  • To address challenges in selecting desired phage genomes.
  • To highlight advances enabling precise phage genetic manipulation.

Main Methods:

  • Discussion of cell-free methodologies for phage genome engineering.
  • Explanation of clustered regularly interspaced short palindromic repeats-CRISPR associated protein (CRISPR-Cas) counter-selection approaches.
  • Integration of these techniques for enhanced phage genome manipulation and selection.

Main Results:

  • Improved methods for understanding and controlling bacteriophage gene functions.
  • Enhanced ability to alter phage properties, such as host range.
  • Facilitation of the development of engineered phages for therapeutic and industrial applications.

Conclusions:

  • Cell-free systems and CRISPR-Cas counter-selection are crucial for advancing bacteriophage engineering.
  • These methods provide universal selectable markers, overcoming previous limitations.
  • The discussed techniques pave the way for novel phage-based solutions in medicine and industry.