Generation of two ISL1-tdTomato reporter human induced pluripotent stem cell lines using CRISPR-Cas9 genome editing

利用 CRISPR-Cas9 基因组编辑生成两种 ISL1-tdTomato 报告人类诱导多能干细胞系

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作者:Satomi Tsukamoto, Koji Nakade, Tamami Wakabayashi, Kenichi Nakashima, Miho Takami, Yasuko Hemmi, Yuji Kuramochi, Tomoya Shimizu, Yutaka Arai, Mami Matsuo-Takasaki, Michiya Noguchi, Yukio Nakamura, Yoshihiro Miwa, Yohei Hayashi

Abstract

ISL1 encodes a member of the LIM/homeodomain family of transcription factors. This encoded protein plays central roles in the development of motor neuron, pancreas, and secondary heart field. Here we generated heterozygous fluorescent reporters of the ISL1 gene in human induced pluripotent stem cells (hiPSCs). CRISPR/Cas9 genome editing technology was employed to knock-in 2A-tdTomato and EF1 alpha promoter-driven Bleomycin resistance gene to the translational ISL1 C-terminal region. The resulting ISL1-TEZ lines showed tdTomato fluorescence upon motor neuron differentiation. These reporter iPSC lines provide opportunity for monitoring and purifying these related cell lineages.

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