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携带沉默抗性钙报告基因等位基因的诱导多能干细胞系。

An induced pluripotent stem cell line carrying a silencing-resistant calcium reporter allele.

机构信息

Nash Family Department of Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA; Institute for Regenerative Medicine, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Institute for Regenerative Medicine, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

出版信息

Stem Cell Res. 2024 Sep;79:103455. doi: 10.1016/j.scr.2024.103455. Epub 2024 Jun 4.

Abstract

Calcium indicators are sensitive tools to image neural activity. However, their use in human induced pluripotent stem cells (iPSC)-derived neurons is limited by silencing of the transgene. We generated the iPSC line MSE2336A carrying heterozygous insertion in the safe-harbor locus AAVS1 of the ultrasensitive protein calcium sensor (GCaMP6) under the control of CAG promoter and UCOE to maintain robust transgene expression in differentiated cells. The iPSC exhibited normal cell morphology, expression of pluripotency markers, genome integrity, and the ability to differentiate into the three primary germ layers. This line provides a powerful model to study activity in human neurons.

摘要

钙指示剂是一种灵敏的工具,可用于对神经活动进行成像。然而,它们在人诱导多能干细胞(iPSC)衍生神经元中的应用受到转基因沉默的限制。我们生成了携带杂合插入的 iPSC 系 MSE2336A,该插入位于 AAVS1 的安全港位点,超灵敏蛋白钙传感器(GCaMP6)由 CAG 启动子和 UCOE 控制,以在分化细胞中维持稳健的转基因表达。iPSC 表现出正常的细胞形态、多能性标记物的表达、基因组完整性以及分化为三个原胚层的能力。该系为研究人类神经元活性提供了一个强大的模型。

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