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Generation of viable PDX1 gene-edited founder pigs as providers of nonmosaics.
Molecular Reproduction and Development ( IF 2.7 ) Pub Date : 2020-03-12 , DOI: 10.1002/mrd.23335 Fuminori Tanihara 1 , Maki Hirata 1 , Nhien Thi Nguyen 1 , Quynh Anh Le 1 , Takayuki Hirano 1 , Takeshige Otoi 1
Molecular Reproduction and Development ( IF 2.7 ) Pub Date : 2020-03-12 , DOI: 10.1002/mrd.23335 Fuminori Tanihara 1 , Maki Hirata 1 , Nhien Thi Nguyen 1 , Quynh Anh Le 1 , Takayuki Hirano 1 , Takeshige Otoi 1
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Pancreatic duodenal homeobox 1 (PDX1) is a crucial gene for pancreas development during the fetal period. PDX1-modified pigs have the potential to be used as a model of diabetes mellitus. However, the severe health problems caused by the PDX1 mutation limit phenotypic studies of PDX1-modified pigs as diabetes models. In this study, we generated PDX1-modified pigs by the CRISPR/Cas9 system introduced into zygotes via electroporation and investigated the mosaicism, phenotypes, and inheritance of the resulting pigs. After the embryo transfer of PDX1-modified zygotes, nine mutant piglets were delivered. Two piglets were apancreatic biallelic mutants. For the other seven piglets, the ratio of mutant alleles to total alleles was 17.5-79.7%. Two mutant piglets with high mutation rates (67.7% and 79.7%) exhibited hypoplasia of the pancreas, whereas the other five piglets were healthy. One of the male mutant piglets was further analyzed. The ejaculated semen from the pig contained PDX1-mutant spermatozoa and the pig showed normal reproductive ability. In conclusion, the frequency of the PDX1 mutation is presumed to relate to pancreas formation, and PDX1 mutant founder pigs generated from zygotes introduced to the CRISPR/Cas9 system can serve as providers of nonmosaics to contribute to medical research on diabetes mellitus.
中文翻译：
生成有活力的PDX1基因编辑的创始猪作为非马赛克提供者。
胰腺十二指肠同源盒1（PDX1）是胎儿期胰腺发育的重要基因。PDX1修饰的猪具有被用作糖尿病模型的潜力。但是，由PDX1突变引起的严重健康问题限制了PDX1修饰猪作为糖尿病模型的表型研究。在这项研究中，我们通过经由电穿孔引入合子的CRISPR / Cas9系统产生了PDX1修饰的猪，并研究了所得猪的镶嵌性，表型和遗传。在PDX1修饰的合子的胚胎转移后，交付了九只突变仔猪。两只小猪是胰腺双等位基因突变体。对于其他七个仔猪，突变等位基因与总等位基因的比例为17.5-79.7％。两只突变率高的突变仔猪（67.7％和79.7％）表现出胰腺发育不全，而其他五头小猪则健康。进一步分析了其中一只雄性突变仔猪。猪的精液中含有PDX1突变型精子，猪的生殖能力正常。总之，PDX1突变的频率被认为与胰腺的形成有关，从CRISPR / Cas9系统引入的受精卵产生的PDX1突变创始人猪可以作为非马赛克提供者，为糖尿病的医学研究做出贡献。
更新日期：2020-03-12
中文翻译：
生成有活力的PDX1基因编辑的创始猪作为非马赛克提供者。
胰腺十二指肠同源盒1（PDX1）是胎儿期胰腺发育的重要基因。PDX1修饰的猪具有被用作糖尿病模型的潜力。但是，由PDX1突变引起的严重健康问题限制了PDX1修饰猪作为糖尿病模型的表型研究。在这项研究中，我们通过经由电穿孔引入合子的CRISPR / Cas9系统产生了PDX1修饰的猪，并研究了所得猪的镶嵌性，表型和遗传。在PDX1修饰的合子的胚胎转移后，交付了九只突变仔猪。两只小猪是胰腺双等位基因突变体。对于其他七个仔猪，突变等位基因与总等位基因的比例为17.5-79.7％。两只突变率高的突变仔猪（67.7％和79.7％）表现出胰腺发育不全，而其他五头小猪则健康。进一步分析了其中一只雄性突变仔猪。猪的精液中含有PDX1突变型精子，猪的生殖能力正常。总之，PDX1突变的频率被认为与胰腺的形成有关，从CRISPR / Cas9系统引入的受精卵产生的PDX1突变创始人猪可以作为非马赛克提供者，为糖尿病的医学研究做出贡献。
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