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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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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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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.
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Related Experiment Video

Updated: Dec 2, 2025

Designing, Packaging, and Delivery of High Titer CRISPR Retro and Lentiviruses via Stereotaxic Injection
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Rapid poxvirus engineering using CRISPR/Cas9 as a selection tool.

Anjali Gowripalan1, Stewart Smith1, Tijana Stefanovic1

  • 1John Curtin School of Medical Research, Australian National University, 131 Garran Road, Acton, ACT, 2601, Australia.

Communications Biology
|November 4, 2020
PubMed
Summary
This summary is machine-generated.

CRISPR/Cas9 gene editing unexpectedly inhibits poxvirus replication by inefficiently repairing viral DNA breaks. This allows for easier selection of engineered poxviruses, accelerating vaccine development for cancer and outbreaks.

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Area of Science:

  • Molecular Biology
  • Virology
  • Gene Editing

Background:

  • CRISPR/Cas9 technology typically relies on efficient DNA repair following genome cutting for genetic modification.
  • Engineering large dsDNA viruses like poxviruses using standard CRISPR/Cas9 methods has been challenging due to repair inefficiencies.

Purpose of the Study:

  • To investigate the efficiency of CRISPR/Cas9-mediated DNA repair in poxviruses.
  • To explore alternative applications of CRISPR/Cas9 in poxvirus genome engineering.

Main Methods:

  • Utilized CRISPR/Cas9 (Cas9 and guide RNA) complexes to target poxvirus genomes post-entry into host cells.
  • Assessed homologous recombination rates between repair constructs and viral genomes.
  • Monitored poxvirus DNA replication and viral spread in cell culture.

Main Results:

  • Cas9-guide RNA complexes effectively cleaved poxvirus genomes but resulted in inefficient repair of the induced breaks.
  • Cas9 cleavage modestly improved, but did not significantly enhance, homologous recombination rates.
  • CRISPR/Cas9 targeting unexpectedly inhibited poxvirus DNA replication and suppressed virus spread.

Conclusions:

  • CRISPR/Cas9 acts as an antiviral mechanism in poxviruses by inhibiting replication, rather than solely facilitating precise genome editing.
  • This inhibitory effect provides a novel method for selecting conventionally generated poxvirus recombinants without marker genes.
  • The application of CRISPR/Cas9 significantly accelerates the generation of poxvirus-based vaccines, enhancing their utility for personalized cancer therapies and rapid response to emerging diseases.