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Generation of Plexin-B1 Conditional Knockout Mouse With CRISPR/Cas9 Technology.

Haofei Ni1,2, Kevin Kelley3, Ning Xie1

  • 1Division of Spine, Department of Orthopedics, Tongji Hospital, Tongji University School of Medicine, Shanghai, China.

Genesis (New York, N.Y. : 2000)
|June 26, 2025
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Summary

Researchers generated a conditional Plexin-B1 (Plxnb1) mouse model using CRISPR/Cas9. This new Plxnb1-floxed allele allows for cell-type specific studies of Plexin-B1 function in development and physiology.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Plexins are crucial axon guidance receptors involved in cytoskeleton and membrane dynamics.
  • Their widespread expression necessitates cell-type specific tools for functional analysis.

Purpose of the Study:

  • To generate a conditional floxed allele of Plexin-B1 (Plxnb1) in mice.
  • To enable future cell type-specific functional studies of Plxnb1.

Main Methods:

  • CRISPR/Cas9 technology was used to insert loxP sites flanking critical exons of the Plxnb1 gene.
  • Targeting reagents were delivered into single-cell mouse embryos via electroporation.
  • PCR and Sanger sequencing were employed for screening and verification.

Main Results:

  • A conditional Plxnb1-floxed allele was successfully generated in mice with a success rate of approximately 1.6%.
  • Alt-R modified ssODNs improved targeting efficiency for one loxP site.
  • The Plxnb1-floxed allele was germline-transmitted and effectively excised using Cre recombinase.

Conclusions:

  • A conditional Plxnb1-floxed mouse model was created using CRISPR/Cas9.
  • This model provides a valuable tool for investigating the specific roles of Plexin-B1 in various biological contexts.