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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

132
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

CRISPR

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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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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and 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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Related Experiment Video

Updated: Aug 19, 2025

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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Advances in Off-Target Detection for CRISPR-Based Genome Editing.

Haozheng Wang1,2,3, Yangmin Wang1,2, Zhongtao Luo1,2

  • 1Guangdong Province Key Laboratory of Biotechnology Drug Candidates, Guangdong Pharmaceutical University, Guangzhou, People's Republic of China.

Human Gene Therapy
|December 1, 2022
PubMed
Summary

CRISPR genome editing holds promise for gene therapy, but off-target effects (OTEs) hinder clinical use. This review details methods for detecting OTEs to ensure safe and effective CRISPR applications.

Keywords:
CRISPR/Cas nucleasesbase editorsgenome editingoff-target detectionprime editors

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas genome editing offers significant potential for gene editing and therapy.
  • Off-target effects (OTEs) remain a major obstacle to the clinical translation of CRISPR technology.
  • Reducing OTEs and developing reliable detection methods are crucial for clinical application.

Purpose of the Study:

  • To summarize existing strategies for detecting OTEs in various CRISPR systems.
  • To provide comprehensive guidance for identifying OTEs in CRISPR-based genome editing applications.
  • To facilitate the safe and effective clinical use of CRISPR technology.

Main Methods:

  • Review and synthesis of current literature on CRISPR OTE detection methods.
  • Categorization of detection strategies based on different CRISPR systems.
  • Analysis of the strengths and limitations of various OTE detection approaches.

Main Results:

  • A comprehensive overview of diverse OTE detection techniques is presented.
  • Guidance is provided for selecting appropriate detection methods for specific CRISPR applications.
  • The importance of unbiased, genome-wide OTE detection is highlighted.

Conclusions:

  • Accurate detection of OTEs is essential for the clinical success of CRISPR genome editing.
  • Continued development of sensitive and unbiased OTE detection methods is necessary.
  • This review serves as a valuable resource for researchers and clinicians working with CRISPR technology.