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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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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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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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Updated: Oct 20, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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Perfecting Targeting in CRISPR.

Hainan Zhang1, Tong Li2, Yidi Sun1

  • 1Institute of Neuroscience, Key Laboratory of Primate Neurobiology, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Center for Brain Science and Brain-Inspired Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China;

Annual Review of Genetics
|September 17, 2021
PubMed
Summary
This summary is machine-generated.

CRISPR gene editing offers great potential for research and therapies. Improving CRISPR specificity is key to reducing unintended DNA or RNA changes, enhancing its use in diverse organisms.

Keywords:
CRISPR-CasCas13RNA editingbase editorgene editingoff-target

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR technology enables precise genome engineering across various organisms.
  • Ensuring the specificity of CRISPR tools is crucial for reliable research and therapeutic outcomes.
  • Off-target effects can limit the application and safety of CRISPR-based genome editing.

Purpose of the Study:

  • To review CRISPR-based DNA and RNA editing technologies.
  • To discuss methods for assessing the specificity of CRISPR editing tools.
  • To highlight strategies for minimizing off-target effects in CRISPR applications.

Main Methods:

  • Literature review of CRISPR-based genome and transcriptome editing technologies.
  • Analysis of methods for quantifying CRISPR specificity (genome-wide and transcriptome-wide).
  • Synthesis of approaches to enhance CRISPR specificity and reduce off-target mutations.

Main Results:

  • Overview of current CRISPR DNA and RNA editing systems.
  • Identification of key techniques for measuring CRISPR tool specificity.
  • Compilation of strategies to improve specificity and mitigate off-target effects.

Conclusions:

  • CRISPR editing technologies are advancing rapidly for diverse applications.
  • Quantifying and enhancing specificity are critical for safe and effective CRISPR use.
  • Minimizing off-target effects will unlock the full therapeutic and research potential of CRISPR systems.