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Related Concept Videos

CRISPR01:59

CRISPR

52.5K
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/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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In-vitro Mutagenesis01:16

In-vitro Mutagenesis

14.1K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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Related Experiment Video

Updated: Aug 4, 2025

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes
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Gene Editing in Mouse Zygotes Using the CRISPR/Cas9 System.

Benedikt Wefers1,2, Wolfgang Wurst3,4,5,6, Ralf Kühn7

  • 1German Center for Neurodegenerative Diseases (DZNE), Munich, Germany. Benedikt.wefers@dzne.de.

Methods in Molecular Biology (Clifton, N.J.)
|March 30, 2023
PubMed
Summary
This summary is machine-generated.

CRISPR/Cas9 gene editing has revolutionized mouse germline engineering, replacing older methods for creating genetically modified mice. This technology enables precise DNA modifications directly in zygotes, accelerating research into gene function.

Keywords:
CRISPRCas9Gene editingGenome engineeringHDRKnockinKnockoutMouseNHEJZygotes

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Area of Science:

  • Biomedical Research
  • Genetics
  • Molecular Biology

Background:

  • Germline engineering in mice is crucial for understanding gene function in health and disease.
  • Traditional gene targeting relied on embryonic stem cell manipulation, a complex and time-consuming process.
  • The advent of CRISPR/Cas9 technology in 2013 marked a significant shift in genetic engineering methodologies.

Purpose of the Study:

  • To detail the application of CRISPR/Cas9 for efficient germline engineering in mice.
  • To provide a comprehensive overview of the gene editing process from guide RNA design to genotyping.
  • To highlight the advantages of CRISPR/Cas9 over previous gene targeting techniques.

Main Methods:

  • Design of guide RNAs (gRNAs) for targeting specific genomic loci.
  • Introduction of Cas9 nuclease and gRNAs into mouse zygotes via microinjection or electroporation.
  • Utilizing DNA repair mechanisms (non-homologous end joining or homology-directed repair) to create gene edits.

Main Results:

  • CRISPR/Cas9 directly induces targeted double-strand breaks in the mouse genome within zygotes.
  • Gene editing results in diverse repair outcomes, including precise knockins or imprecise knockouts.
  • This method has become the standard for generating genetically engineered mice due to its efficiency and ease of use.

Conclusions:

  • CRISPR/Cas9 technology offers a rapid and effective means for generating genetically engineered mice.
  • The ease of application in zygotes has democratized mouse germline engineering for biomedical research.
  • This review covers essential aspects of CRISPR/Cas9-mediated gene editing in mice, facilitating future research.