title: 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.
authors.affiliation: Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, India.
authors.forename: Deeti K
authors.initials: DK
authors.lastname: Shetty
authors.affiliation: Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru, India; Institute for Stem Cell Biology and Regenerative Medicine, Bengaluru, India.
authors.forename: Maneesha S
authors.initials: MS
authors.lastname: Inamdar
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D058732:Embryoid Bodies / Q000166:cytology; D044783:F-Box Proteins / Q000235:genetics / Q000378:metabolism*; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D059785:Karyotype; D051379:Mice; D008856:Microscopy, Fluorescence; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: C509434:Emi2 protein, mouse; D044783:F-Box Proteins; D014157:Transcription Factors