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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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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CRISPR01:59

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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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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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Related Experiment Video

Updated: Apr 19, 2026

Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing
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Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing

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Targeting Hepatitis B Virus With CRISPR/Cas9.

Christoph Seeger1, Ji A Sohn1

  • 1Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA.

Molecular Therapy. Nucleic Acids
|December 17, 2014
PubMed
Summary

The CRISPR/Cas9 system effectively targets and disrupts Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) in liver cells. This breakthrough offers a potential new strategy for eliminating persistent HBV infections by destroying the viral DNA template.

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CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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Area of Science:

  • Hepatology
  • Molecular Virology
  • Gene Editing Technologies

Background:

  • Hepatitis B virus (HBV) persistence relies on nuclear covalently closed circular DNA (cccDNA), a target resistant to current therapies.
  • Nucleoside analogues inhibit HBV replication but do not eliminate the cccDNA template.

Purpose of the Study:

  • To investigate the efficacy of the CRISPR/Cas9 system in directly targeting and cleaving HBV cccDNA.
  • To assess the potential of gene editing as a novel antiviral strategy against persistent HBV infection.

Main Methods:

  • Utilized the CRISPR/Cas9 gene editing system in HepG2 cells engineered to express the HBV receptor NTCP.
  • Designed and tested HBV-specific guide RNAs to direct Cas9 cleavage of cccDNA.
  • Analyzed cccDNA integrity and HBV inhibition post-CRISPR/Cas9 treatment.

Main Results:

  • CRISPR/Cas9 targeting significantly inhibited HBV infection, reducing viral activity up to eightfold.
  • Inhibition resulted from Cas9-induced mutations and deletions in cccDNA, repaired via nonhomologous end joining (NHEJ).
  • Interferon alpha (IFN-α) did not impede CRISPR/Cas9 antiviral activity, indicating robustness against innate immune responses.

Conclusions:

  • Cas9 can be effectively recruited to HBV cccDNA for endonucleolytic cleavage.
  • CRISPR/Cas9 demonstrates potential for developing novel antiviral therapies aimed at eradicating persistent HBV by destroying cccDNA.