Objective To breed and identify NLRP3 gene knock-out mice. Methods The NLRP3 gene knock-out heterozygote mice were bred alone and copulated. The offsprings were to have three genotypes: wild genotype, heterozygote genotype and homozygote genotype. Genomic DNA was obtained from each pups and were subjected to PCR and T7 endonuclease 1 to identify the genotype. The homozygote mice were mated with the opposite sex heterozygote mice to obtain more homozygote pups. Results Breeding and reproducing were both successful, and we obtained heterozygote genotype and homozygote genotype mice with NLRP3 gene knock-out. Conclusion Correct methods of breeding, reproducing and identifying can effectively obtain NLRP3 gene knock-out mice from heterozygote mice.