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Related Concept Videos

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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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Related Experiment Video

Updated: May 11, 2025

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Genome Editing in Mouse Embryo Using the CRISPR/Cas12i3 System.

Jiale He1, Juan Liu1, Yuan Yue1

  • 1Frontiers Science Center for Molecular Design Breeding (MOE), Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture and Rural Affairs, National Engineering Laboratory for Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.

International Journal of Molecular Sciences
|April 17, 2025
PubMed
Summary

The CRISPR/Cas12i3 gene editing tool shows high efficiency and safety in mouse embryos, successfully creating a Nanog loss-of-function model. This system offers a reliable method for generating gene-edited animals for research and breeding.

Keywords:
CRISPR/Cas12i3Nanoggenome editingmouse embryo

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Area of Science:

  • Molecular Biology
  • Genetics
  • Developmental Biology

Background:

  • CRISPR/Cas systems are advanced gene editing tools.
  • Cas12i3 offers unique features like T-rich PAM recognition and high efficiency in rice.
  • Previous applications of Cas12i3 have not been reported in mammalian embryos.

Purpose of the Study:

  • To evaluate the feasibility and safety of CRISPR/Cas12i3 for gene editing in mouse embryos.
  • To target the core pluripotency regulator gene Nanog.
  • To establish a Nanog loss-of-function mouse embryo model.

Main Methods:

  • Systematic evaluation of CRISPR/Cas12i3 in mouse embryos.
  • Targeting the Nanog gene locus.
  • Optimization of Cas12i3 mixture concentration.

Main Results:

  • CRISPR/Cas12i3 achieved high editing efficiency at the Nanog locus, surpassing Cas9.
  • No off-target mutations were detected.
  • Cas12i3 demonstrated no adverse effects on mouse embryo development or blastocyst cell proliferation.
  • Healthy chimeric gene-edited offspring were produced.

Conclusions:

  • CRISPR/Cas12i3 is a feasible and safe gene editing tool for mammalian embryos.
  • This system provides a reliable method for one-step generation of gene-edited animals.
  • Applications include biology, medical research, and livestock breeding.