Construction and disease research of CXCR4 gene knockout mouse model

Objective: By using CRISPR/Cas9 technology, we generated the C-X-C Motif Chemokine Receptor 4 (CXCR4) gene-knockout mouse via electroporation of fertilized eggs, so as to elucidate the function of CXCR4 diseases, and provide transgenic animal resources for future disease modeling and therapeutic research. Methods: Superovulation and fertilized egg collection were performed, followed by plasmid vector construction, primer design, vector preparation, transcription in vitro, and pronuclear injection. Strictly, mixture of Cas9 mRNA and sgRNA was microinjected into fertilized eggs, which were then transferred to pseudopregnant recipient female mice. Genotyping of the F0 generation was performed through breeding and confirmed by PCR and sequencing. Results: The CXCR4 gene-knockout mice model was successfully constructed using CRISPR/Cas9 technology. PCR and sequencing confirmed the gene knockout in the F1 generation. Among 23 mice genotyped, 8 were positive for CXCR4 knockout, while the remainder were wild-type, validating the success of the knockout process for further research. Conclusions: We established the CXCR4 gene-knockout mice model, which is valuable for further investigations into the function of the CXCR4 gene and its involvement in various diseases.