Romero-Moya Damià, Castaño Julio, Santos-Ocaña Carlos, Navas Plácido, Menendez Pablo
Josep Carreras Leukemia Research Institute, Department of Biomedicine, School of Medicine, University of Barcelona, Barcelona, Spain.
Josep Carreras Leukemia Research Institute, Department of Biomedicine, School of Medicine, University of Barcelona, Barcelona, Spain.
Stem Cell Res. 2017 Oct;24:144-147. doi: 10.1016/j.scr.2016.09.007. Epub 2016 Sep 16.
We report the generation, CRISPR/Cas9-edition and characterization of induced pluripotent stem cell (iPSC) lines from a patient with coenzyme Q deficiency harboring the heterozygous mutation c.483G>C in the COQ4 gene. iPSCs were generated using non-integrative Sendai Viruses containing the reprogramming factors OCT4, SOX2, KLF4 and C-MYC. The iPSC lines carried the c.483G>C COQ4 mutation, silenced the OKSM expression and were mycoplasma-free. They were bona fide pluripotent cells as characterized by morphology, immunophenotype/gene expression for pluripotent-associated markers/genes, NANOG and OCT4 promoter demethylation, karyotype and teratoma formation. The COQ4 mutation was CRISPR/Cas9 edited resulting in isogenic, diploid and off-target free COQ4-corrected iPSCs.
我们报告了从一名患有辅酶Q缺乏症且COQ4基因存在杂合突变c.483G>C的患者中诱导多能干细胞（iPSC）系的产生、CRISPR/Cas9编辑及特性鉴定。使用含有重编程因子OCT4、SOX2、KLF4和C-MYC的非整合仙台病毒来产生iPSC。这些iPSC系携带c.483G>C的COQ4突变，沉默了OKSM表达且无支原体污染。通过形态学、多能性相关标志物/基因的免疫表型/基因表达、NANOG和OCT4启动子去甲基化、核型及畸胎瘤形成鉴定，它们是真正的多能干细胞。对COQ4突变进行了CRISPR/Cas9编辑，得到了等基因、二倍体且无脱靶效应的COQ4校正iPSC。
