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Updated: Oct 8, 2025
CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions
Published on: November 1, 2024
Generation of a B2M homozygous knockout human somatic cell nuclear transfer-derived embryonic stem cell line using
Ok-Hee Lee1, Siyoung Lee2, Miseon Park3
1Department of Biomedical Science, CHA University, Seongnam-si, Gyeonggi-do 13488, Republic of Korea; Department of Veterinary Physiology, College of Veterinary Medicine, Konkuk University, Seoul 05029, Republic of Korea.
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.
