利用 CRISPR-Cas9 系统在胰岛素启动子区域内对 GG2-GG1/A2 元件活性进行体内评估。

In vivo evaluation of GG2-GG1/A2 element activity in the insulin promoter region using the CRISPR-Cas9 system.

机构信息

Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Nishihara, Okinawa, 903-0215, Japan.

Department of Basic Laboratory Sciences, School of Health Sciences in Faculty of Medicine, University of the Ryukyus, Nishihara, Okinawa, 903-0215, Japan.

出版信息

Sci Rep. 2021 Oct 13;11(1):20290. doi: 10.1038/s41598-021-99808-6.

DOI:10.1038/s41598-021-99808-6

PMID:34645928

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8514523/

Abstract

The insulin promoter is regulated by ubiquitous as well as pancreatic β-cell-specific transcription factors. In the insulin promoter, GG2-GG1/A2-C1 (bases - 149 to - 116 in the human insulin promoter) play important roles in regulating β-cell-specific expression of the insulin gene. However, these events were identified through in vitro studies, and we are unaware of comparable in vivo studies. In this study, we evaluated the activity of GG2-GG1/A2 elements in the insulin promoter region in vivo. We generated homozygous mice with mutations in the GG2-GG1/A2 elements in each of the Ins1 and Ins2 promoters by CRISPR-Cas9 technology. The mice with homozygous mutations in the GG2-GG1/A2 elements in both Ins1 and Ins2 were diabetic. These data suggest that the GG2-GG1/A2 element in mice is important for Ins transcription in vivo.

摘要

胰岛素启动子受普遍存在的以及胰腺β细胞特异性转录因子调节。在胰岛素启动子中，GG2-GG1/A2-C1（人胰岛素启动子中的-149 到-116 位碱基）在调节胰岛素基因的β细胞特异性表达中发挥重要作用。然而，这些事件是通过体外研究确定的，我们还不知道有类似的体内研究。在这项研究中，我们评估了胰岛素启动子区域 GG2-GG1/A2 元件在体内的活性。我们通过 CRISPR-Cas9 技术在 Ins1 和 Ins2 启动子中的 GG2-GG1/A2 元件中生成了纯合突变的小鼠。Ins1 和 Ins2 中 GG2-GG1/A2 元件的纯合突变小鼠患有糖尿病。这些数据表明，在体内，小鼠的 GG2-GG1/A2 元件对于 Ins 转录是重要的。