Knockout of Tyr gene by CRISPR-Cas9 to produce albino C57BL / 6N mice
Received:February 14, 2019  
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DOI:10. 3969 / j.issn.1005-4847. 2019. 04. 011
KeyWord:Albino; Tyr; gene knockout; CRISPR-Cas9; C57BL/6N; mouse
                       
AuthorInstitution
常秋荣 上海交通大学医学院实验动物科学部,上海 
刘丽丽 上海交通大学医学院实验动物科学部,上海 
王会阳 上海交通大学医学院实验动物科学部,上海 
付丽 上海交通大学医学院实验动物科学部,上海 
邢凤英 上海交通大学医学院实验动物科学部,上海 
李垚 上海交通大学医学院实验动物科学部,上海 
陈学进 上海交通大学医学院实验动物科学部,上海 
李善刚 1. 上海交通大学医学院实验动物科学部,上海 ; 2. 昆明理工大学灵长类转化医学研究院,昆明 
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Abstract:
      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.
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