Mutations in the C1 element of the insulin promoter lead to diabetic phenotypes in homozygous mice
Mutations in the C1 element of the insulin promoter lead to diabetic phenotypes in homozygous mice
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DOI：
10.1038/s42003-020-1040-z
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发表时间：
2020-06-16
影响因子：
5.9
通讯作者：
Watanabe, Masami
中科院分区：
文献类型：
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作者：
Noguchi, Hirofumi;Miyagi-Shiohira, Chika;Watanabe, Masami
CRISPR-Cas9等基因组编辑技术被广泛用于建立突变和表型之间的因果关系。然而，CRISPR-Cas9很少用于分析启动子区域。胰岛素启动子区（约1,000bp）指导β细胞特异性胰岛素表达，体外研究表明，胰岛素表达受普遍存在的β细胞特异性转录因子以及胰腺特异性转录因子的调节。然而，我们不知道任何证实的体内研究。在这里，我们使用CRISPR-Cas9技术产生了胰岛素I （Ins1）和II （Ins2）基因启动子区域突变的小鼠。我们培育了4只纯合子糖尿病小鼠，在Ins1和Ins2启动子中高度保守的C1元件各有2个不同的突变（总共有3个缺失和1个替换）。值得注意的是，所有在其他基因座有纯合或杂合突变的小鼠都没有患糖尿病。因此，小鼠体内的Ins转录需要C1元件。Noguchi等人利用CRISPR-Cas9产生了胰岛素I和II基因启动子区域突变的小鼠，并表明只有高度保守的C1元件的纯合突变触发糖尿病表型，突出了该基因座在胰岛素转录中的重要性。
Genome editing technologies such as CRISPR-Cas9 are widely used to establish causal associations between mutations and phenotypes. However, CRISPR-Cas9 is rarely used to analyze promoter regions. The insulin promoter region (approximately 1,000bp) directs beta cell-specific expression of insulin, which in vitro studies show is regulated by ubiquitous, as well as pancreatic, beta cell-specific transcription factors. However, we are unaware of any confirmatory in vivo studies. Here, we used CRISPR-Cas9 technology to generate mice with mutations in the promoter regions of the insulin I (Ins1) and II (Ins2) genes. We generated 4 homozygous diabetic mice with 2 distinct mutations in the highly conserved C1 elements in each of the Ins1 and Ins2 promoters (3 deletions and 1 replacement in total). Remarkably, all mice with homozygous or heterozygous mutations in other loci were not diabetic. Thus, the C1 element in mice is required for Ins transcription in vivo. Noguchi et al. use CRISPR-Cas9 to generate mice with mutations in the promoter regions of the insulin I and II genes and show that only homozygous mutations in the highly conserved C1 element triggered a diabetic phenotype, highlighting the importance of this loci in insulin transcription.