α-Gal antigen-deficient rabbits with GGTA1 gene disruption via CRISPR/Cas9

Autor: Lina Wei, Yufeng Mu, Jichao Deng, Yong Wu, Ying Qiao, Kun Zhang, Xuewen Wang, Wenpeng Huang, Anliang Shao, Liang Chen, Yang Zhang, Zhanjun Li, Liangxue Lai, Shuxin Qu, Liming Xu
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: BMC Genomic Data, Vol 23, Iss 1, Pp 1-10 (2022)
Druh dokumentu: article
ISSN: 2730-6844
DOI: 10.1186/s12863-022-01068-4
Popis: Abstract Background Previous studies have identified the carbohydrate epitope Galα1–3Galβ1–4GlcNAc-R (termed the α-galactosyl epitope), known as the α-Gal antigen as the primary xenoantigen recognized by the human immune system. The α-Gal antigen is regulated by galactosyltransferase (GGTA1), and α-Gal antigen-deficient mice have been widely used in xenoimmunological studies, as well as for the immunogenic risk evaluation of animal-derived medical devices. The objective of this study was to develop α-Gal antigen-deficient rabbits by GGTA1 gene editing with the CRISPR/Cas9 system. Results The mutation efficiency of GGTA1 gene-editing in rabbits was as high as 92.3% in F0 pups. Phenotype analysis showed that the α-Gal antigen expression in the major organs of F0 rabbits was decreased by more than 99.96% compared with that in wild-type (WT) rabbits, and the specific anti-Gal IgG and IgM antibody levels in F1 rabbits increased with increasing age, peaking at approximately 5 or 6 months. Further study showed that GGTA1 gene expression in F2-edited rabbits was dramatically reduced compared to that in WT rabbits. Conclusions α-Gal antigen-deficient rabbits were successfully generated by GGTA1 gene editing via the CRISPR/Cas9 system in this study. The feasibility of using these α-Gal antigen-deficient rabbits for the in situ implantation and residual immunogenic risk evaluation of animal tissue-derived medical devices was also preliminarily confirmed.
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