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一种异位表达内体蛋白Asrij(该蛋白调节小鼠胚胎干细胞的多能性)的转基因人类胚胎干细胞系的生成:BJNhem20-Asrij。

Generation of a transgenic human embryonic stem cell line ectopically expressing the endosomal protein Asrij that regulates pluripotency in mouse embryonic stem cells: BJNhem20-Asrij.

作者信息

Shetty Deeti K, Inamdar Maneesha S

机构信息

Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, India.

Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, India; Institute for Stem Cell Biology and Regenerative Medicine, Bengaluru, India.

出版信息

Stem Cell Res. 2016 Mar;16(2):331-3. doi: 10.1016/j.scr.2015.12.044. Epub 2016 Jan 6.

Abstract

Asrij is an endocytic protein expressed in mouse embryonic stem cells (mESCs) and is essential for maintenance of stemness of mESCs (Mukhopadhyay et al., 2003; Sinha et al., 2013). Its human ortholog named Ovarian Carcinoma Immuno-reactive Antigen domain containing protein 1 (OCIAD1) is 85% identical. We ectopically expressed Asrij in epiblast stage equivalent-human embryonic stem cells (hESCs) to test for induction of naive pluripotency in primed pluripotent cells. The construct pCAG-Asrij was introduced into hESCs by microporation. Ectopic expression of Asrij in BJNhem20 hESC line was performed by selecting for plasmid transfection, followed by stable cell line generation.

摘要

Asrij是一种在小鼠胚胎干细胞(mESCs)中表达的内吞蛋白,对维持mESCs的干性至关重要(Mukhopadhyay等人,2003年;Sinha等人,2013年)。其人类同源物名为含卵巢癌免疫反应抗原结构域蛋白1(OCIAD1),两者有85%的同源性。我们在相当于上胚层阶段的人类胚胎干细胞(hESCs)中异位表达Asrij,以测试其对已分化多能细胞诱导原始多能性的作用。通过微穿孔将构建体pCAG-Asrij导入hESCs。通过选择质粒转染,然后生成稳定细胞系,在BJNhem20 hESC系中进行Asrij的异位表达。

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