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

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

doi:10.1016/j.scr.2021.102643

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

Stem Cell Res. 2022 Mar:59:102643. doi: 10.1016/j.scr.2021.102643. Epub 2021 Dec 27.

Authors

Ok-Hee Lee¹, Siyoung Lee², Miseon Park³, Sohyeon Moon², Semi Hwang², Byeongseok Kim², C-Yoon Kim⁴, Dong Ryul Lee⁵, Sung Han Shim⁵, Keun-Hong Park⁵, Hyung Min Chung⁶, Youngsok Choi⁷

Affiliations

¹ Department 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.
² Department of Stem Cell and Regenerative Biotechnology, Konkuk University, Seoul 05029, Republic of Korea.
³ Fertility Center of CHA Gangnam Medical Center, Department of Obstetrics and Gynecology, CHA University, Seoul 06135, Republic of Korea.
⁴ Department of Veterinary Physiology, College of Veterinary Medicine, Konkuk University, Seoul 05029, Republic of Korea.
⁵ Department of Biomedical Science, CHA University, Seongnam-si, Gyeonggi-do 13488, Republic of Korea.
⁶ Department of Stem Cell Biology, School of Medicine, Konkuk University, Seoul 05029, Republic of Korea.
⁷ Department of Stem Cell and Regenerative Biotechnology, Konkuk University, Seoul 05029, Republic of Korea. Electronic address: choiys3969@konkuk.ac.kr.

PMID: 34971932
DOI: 10.1016/j.scr.2021.102643

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.