Balboa Diego, Weltner Jere, Novik Yuval, Eurola Solja, Wartiovaara Kirmo, Otonkoski Timo
Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland.
Research Programs Unit, Molecular Neurology and Biomedicum Stem Cell Centre, Faculty of Medicine, University of Helsinki, 00014 Helsinki, Finland.
Stem Cell Res. 2017 Jul;22:16-19. doi: 10.1016/j.scr.2017.05.005. Epub 2017 May 17.
SOX2 is an important transcription factor involved in pluripotency maintenance, pluripotent reprogramming and differentiation towards neural lineages. Here we engineered the previously described HEL24.3 hiPSC to generate a SOX2 reporter by knocking-in a T2A fused nuclear tdTomato reporter cassette before the STOP codon of the SOX2 gene coding sequence. CRISPR/SaCas9-mediated stimulation of homologous recombination was utilized to facilitate faithful targeted insertion. This line accurately reports the expression of endogenous SOX2 and therefore constitutes a useful tool to study the SOX2 expression dynamics upon hiPSC culture, differentiation and somatic cell reprogramming.
SOX2是一种重要的转录因子，参与多能性维持、多能性重编程以及向神经谱系的分化。在此，我们对先前描述的HEL24.3人诱导多能干细胞（hiPSC）进行改造，通过在SOX2基因编码序列的终止密码子之前敲入一个与核tdTomato报告基因盒融合的T2A，来生成一个SOX2报告基因。利用CRISPR/SaCas9介导的同源重组刺激来促进准确的靶向插入。该细胞系能够准确报告内源性SOX2的表达，因此构成了一个有用的工具，用于研究hiPSC培养、分化和体细胞重编程过程中SOX2的表达动态。
