Shetty Deeti K, Inamdar Maneesha S
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.
Stem Cell Res. 2016 Mar;16(2):331-3. doi: 10.1016/j.scr.2015.12.044. Epub 2016 Jan 6.
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.
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。
