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Related Concept Videos

CRISPR01:59

CRISPR

46.2K
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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Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes
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Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

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A mouse geneticist's practical guide to CRISPR applications.

Priti Singh1, John C Schimenti2, Ewelina Bolcun-Filas1

  • 1Department of Biomedical Sciences, Cornell University, Ithaca, New York 14850.

Genetics
|October 2, 2014
PubMed
Summary

The CRISPR/Cas9 system offers a fast and simple way to edit mouse genomes, enabling the creation of various genetic modifications. Optimizations focus on improving efficiency and addressing challenges like mosaicism for better research outcomes.

Keywords:
CRISPRgenome editingmouse knockoutsnonhomologous end joining

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Area of Science:

  • Genetics
  • Molecular Biology
  • Bioengineering

Background:

  • The CRISPR/Cas9 system is a powerful RNA-guided genome editing tool.
  • It revolutionizes genetic research across many organisms, including the laboratory mouse.
  • CRISPR/Cas9 offers unprecedented simplicity and speed for genome manipulation in mice.

Purpose of the Study:

  • To provide a practical guide for using CRISPR/Cas9 for mouse mutagenesis.
  • To summarize current applications and suggest technical improvements for genome editing in mouse embryos.
  • To address practical challenges such as mosaicism and improve genotyping/phenotyping.

Main Methods:

  • Application of CRISPR/Cas9 technology for generating diverse mouse alleles (null, conditional, mutated, reporter, tagged).
  • Development of strategies to increase efficiency of RNA-guided genome editing in mouse embryos.
  • Utilizing next-generation sequencing for on- and off-target editing characterization.
  • Transient inhibition of the nonhomologous end-joining pathway in one-celled mouse embryos.

Main Results:

  • CRISPR/Cas9 enables rapid generation of various precisely engineered mouse models.
  • Technical improvements are proposed to enhance genome editing efficiency and reduce mosaicism.
  • A next-generation sequencing method is presented for comprehensive editing analysis.
  • Inhibition of nonhomologous end-joining increases homology-directed editing frequency.

Conclusions:

  • CRISPR/Cas9 technology significantly advances mouse genetics research.
  • Optimized protocols and analytical strategies are crucial for efficient and accurate genome editing.
  • Targeted inhibition of DNA repair pathways can enhance precise genome editing outcomes in mouse embryos.