Generation of a SOX2 reporter human induced pluripotent stem cell line using CRISPR/SaCas9

利用 CRISPR/SaCas9 生成 SOX2 报告人类诱导多能干细胞系

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作者:Diego Balboa, Jere Weltner, Yuval Novik, Solja Eurola, Kirmo Wartiovaara, Timo Otonkoski

Abstract

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.

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