The objective of this study was to disrupt the low density lipoprotein receptor gene(LDLR) of pig cells using the CRISPR/Cas9 system. Specific guide RNA(sgRNA) sequences were designed based on the LDLR sequence from the NCBI database(Gene ID 396801). A CRISPR/Cas9 target vector was constructed and porcine embryonic fibroblasts colonies were generated through electrotransfection and G418 screening. Genotype identification was performed on the cell colonies. PCR products were cloned into a T vector and subjected to sequence alignment. The efficiency of gene knockout was analyzed statistically. The sgRNA was designed to target the first exon of the LDLR following the GN20GG rule. DNA sequence assays confirmed the correct construction of the target vector, which was subsequently introduced into porcine fetal fibroblasts using electroporation. In total, 30 monoclonal cells were isolated, with successful sequencing of 23 colonies. Among these, seven cell colonies exhibited gene modifications, resulting in a fragment knockout efficiency of 30.4%.