The sgRNA site was designed by the chopchop website on the second exon of the rat Inhba gene and was constructed into the px330 vector by synthesizing sgRNA oligonucleotides and enzyme digestion and ligation. The px330-Inhba-sgRNA vector was transfected into rat L6 cells, and the efficiency of sgRNA on Inhba gene editing was verified by T7E1 digestion and T vector cloning and sequencing. The results showed that after transfection of the pX330-Inbha-sgRNA vector, the PCR product in the editing region of the Inhba gene could be cleaved by the T7E1 enzyme to produce the expected fragment. T vector cloning and sequencing showed that five in the selected randomly twenty clones showed different lengths of base deletion at the targeted cutting sites, and the estimated editing efficiency was 25%. In summary, it showed that the sgRNA designed in this study could effectively edit the Inhba gene using the CRISPR/Cas9 system in rat.