Objective To obtain albino C57BL/6N mice and expand their application in research of skintransplantation and embryonic stem cells. Methods Cas9 mRNA and a series of single guide RNAs (sgRNAs) weresynthesized in vitro and injected into the fertilized eggs of C57BL/6N mice. The gene encoding tyrosinase (TYR), anenzyme necessary for melanin production in C57BL/6N mice, was destroyed in exon 1 and exon 2, and gene mutations weregenerated to obtain albino F0 generation. Albino C57BL/6N mice were then subjected to repeated backcrossing andinbreeding to produce C57BL/6N albino mouse inbreds. Results Two pairs of sgRNA were injected into mice, and the F0generation of albino mice was successfully obtained. Deletions in both exon 1 and exon 2 of the Tyr gene were confirmed.The albino mice transmited the mutant gene to offsprings, and homozygous white C57BL/6N mice as offspring wereconfirmed. The mutation types of albino mice were analyzed. Conclusions The mouse Tyr gene is disrupted by CRISPRCas9technology, and the C57BL/6N albino mouse inbred line is successfully established. This mouse line provides a new research tool for future chimera preparation and tissue transplantation.