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Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9
Published on: January 3, 2015
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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
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.
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.
