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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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RNA viruses are categorized into positive-strand, negative-strand, or double-stranded groups based on their genomic structure and replication mechanisms. This classification dictates how they exploit host cellular machinery for protein synthesis and replication. Some RNA viruses also utilize reverse transcription as part of their life cycle, further diversifying their replication strategies.Positive-Strand RNA VirusesPositive-strand RNA viruses have genomes that function directly as messenger...
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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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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.
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Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the...
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Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
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First Versatile Reverse Genetics System for DNA Viruses Using Circular Polymerase Extension Reaction.

Hirotaka Yamamoto1, Tomokazu Tamura1,2,3,4,5, Rigel Suzuki1,2,4

  • 1Department of Microbiology and Immunology, Faculty of Medicine, Hokkaido University, Sapporo, Japan.

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This study introduces circular polymerase extension reaction (CPER) for rapid DNA virus reverse genetics. The method efficiently generates infectious recombinant adenoviruses (AdVs), accelerating AdV research and therapeutic development.

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DNA virusadenoviruscircular polymerase extension reactionreverse genetics

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Reverse genetics is crucial for virus research but conventional methods for DNA viruses are complex.
  • Circular polymerase extension reaction (CPER) is established for RNA viruses but not DNA viruses.

Purpose of the Study:

  • To adapt and apply CPER for efficient generation of infectious recombinant DNA viruses.
  • To evaluate CPER as a novel reverse genetics platform for adenoviruses (AdVs).

Main Methods:

  • Applied CPER to generate full-length cDNA of two adenovirus serotypes.
  • Assessed the infectivity and replication of generated recombinant AdVs.
  • Confirmed viral replication using immunostaining techniques.

Main Results:

  • Successfully generated infectious recombinant adenoviruses (AdVs) using CPER.
  • Recombinant AdVs exhibited replication comparable to parental strains.
  • CPER proved to be a rapid and efficient method for DNA virus reverse genetics.

Conclusions:

  • CPER is a viable and efficient platform for reverse genetics of DNA viruses.
  • This method can accelerate adenovirus research and the development of AdV-based therapeutics.
  • The findings open new avenues for genetic manipulation of DNA viruses.