Generation of a B2M homozygous knockout human somatic cell nuclear transfer-derived embryonic stem cell line using the CRISPR/Cas9 system

利用 CRISPR/Cas9 系统生成 B2M 纯合敲除人类体细胞核移植衍生的胚胎干细胞系

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作者:Ok-Hee Lee, Siyoung Lee, Miseon Park, Sohyeon Moon, Semi Hwang, Byeongseok Kim, C-Yoon Kim, Dong Ryul Lee, Sung Han Shim, Keun-Hong Park, Hyung Min Chung, Youngsok Choi

Abstract

Beta2-microglobulin (B2M) is a subunit of human leukocyte antigen class-I (HLA-I) heterodimer that mediates immune rejection through activation of cytotoxic T cells. B2M binding to HLA-I proteins is essential for functional HLA-I on the cell surface. Here, we generated a B2M homozygous knockout somatic cell nuclear transfer-induced embryonic stem cell (SCNT-ESC) line using CRISPR/Cas9-mediated gene targeting. B2M KO cell line, which does not express HLA-I molecules on cell surface, has pluripotency and differentiation ability to three germ layers. This cell line provides a useful cell source for investigating immunogenicity of allogeneic ESCs and their derivatives for tissue regeneration.

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