Generation of hiPSCs with ABO c.767T>C substitution: resulting in splicing variants

Yinge Jin; Tao Chen; Wei Zheng; Jiahui Xi; Yin Zi; Jinling Wang; Yue Chi; Min Chen; Qingjian Zou; Chengcheng Tang; Liangxue Lai; Xiaoqing Zhou

doi:10.3389/fgene.2023.1141756

Generation of hiPSCs with ABO c.767T>C substitution: resulting in splicing variants

Front Genet. 2023 Jun 15:14:1141756. doi: 10.3389/fgene.2023.1141756. eCollection 2023.

Authors

Yinge Jin¹, Tao Chen¹, Wei Zheng¹, Jiahui Xi¹, Yin Zi¹, Jinling Wang¹, Yue Chi¹, Min Chen¹, Qingjian Zou¹, Chengcheng Tang¹, Liangxue Lai^{1

2}, Xiaoqing Zhou¹

Affiliations

¹ Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Science, South China Institute of Large Animal Models for Biomedicine, Wuyi University, Jiangmen, China.
² Chinese Academy of Sciences, Guangzhou Institutes of Biomedicine and Health, Guangzhou, China.

Abstract

Introduction: The ABO blood group system has important clinical significance in the safety of blood transfusion and organ transplantation. Numerous ABO variations, especially variations in the splice sites, have been identified to be associated with some ABO subtypes. Methods: Here, we performed the c.767T>C substitution of the ABO gene in human induced pluripotent stem cells (hiPSCs) by the adenosine base editor (ABE) system and described its characteristics at the genome level in detail. Results: The hiPS cell line with c.767T>C substitution maintained a normal karyotype (46, XX), expressed pluripotency markers, and showed the capability to spontaneously differentiate into all three germ layers in vivo. The genome-wide analysis demonstrated that the c.767T>C substitution in the ABO gene did not cause any detected negative effect in hiPSCs at the genome level. The splicing transcript analysis revealed that splicing variants were observed in the hiPSCs with ABO c.767T>C substitutions. Conclusion: All these results indicated that some splicing variants occurred in hiPSCs with c.767 T>C substitution of ABO gene, which probably had a significant effect on the formation of the rare ABO*Ael05/B101 subtype.

Keywords: ABO gene; base substitution; hiPSC; rare subtype; splicing variants.

Grants and funding

This research was funded by the Youth Innovation Project of Guangdong Province University (2022KQNCX095), the Science and Technology Planning Project of Jiangmen (2021030101220004887 and 2021030101230004833), and the Natural Science Foundation of Guangdong Province of China (2019A1515110283).