TY - JOUR
T1 - Mutations in the C1 element of the insulin promoter lead to diabetic phenotypes in homozygous mice
AU - Noguchi, Hirofumi
AU - Miyagi-Shiohira, Chika
AU - Nakashima, Yoshiki
AU - Kinjo, Takao
AU - Saitoh, Issei
AU - Watanabe, Masami
N1 - Funding Information:
We thank Naomi Kakazu (University of the Ryukyus) for administrative support and Maki Higa, Yuki Kawahira, Ikue Honda, Yuka Onishi, Saori Adaniya, Youichi Toyokawa (University of the Ryukyus), Seiya Mizuno, Yoko Tanimoto, Satoru Takahashi, and Fumihiro Sugiyama (University of Tsukuba) for technical support. This work was supported in part by JSPS KAKENHI Grant Numbers JP17H00769 and JP19K09051 and Okinawa Science and Technology Innovation System Construction Project.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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 1,000 bp) directs β cell-specific expression of insulin, which in vitro studies show is regulated by ubiquitous, as well as pancreatic, β 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.
AB - 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 1,000 bp) directs β cell-specific expression of insulin, which in vitro studies show is regulated by ubiquitous, as well as pancreatic, β 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.
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U2 - 10.1038/s42003-020-1040-z
DO - 10.1038/s42003-020-1040-z
M3 - Article
C2 - 32546815
AN - SCOPUS:85086583946
SN - 2399-3642
VL - 3
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 309
ER -
