iPS Cell Advanced Characterization and Development Team, BioResource Research Center, RIKEN, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.
Gene Engineering Division, BioResource Research Center, RIKEN, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.
Stem Cell Res. 2021 May;53:102363. doi: 10.1016/j.scr.2021.102363. Epub 2021 Apr 22.
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.
ISL1 编码 LIM/homeodomain 转录因子家族的成员。该编码蛋白在运动神经元、胰腺和次级心脏场的发育中发挥核心作用。在这里，我们在人诱导多能干细胞（hiPSC）中生成了 ISL1 基因的杂合荧光报告基因。CRISPR/Cas9 基因组编辑技术被用于将 2A-tdTomato 和 EF1 alpha 启动子驱动的博来霉素抗性基因敲入到 ISL1 翻译 C 末端区域。所得的 ISL1-TEZ 系在神经元分化时显示 tdTomato 荧光。这些报告基因 iPSC 系为监测和纯化这些相关细胞谱系提供了机会。
