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

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.