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| 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 |
| Author | Institution |
| 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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