Building method of donor pig suitable for xenotransplantation of tissue like islet cells and skin

Abstract

The invention discloses a building method of a donor pig suitable for xenotransplantation of tissue like islet cells and skin and belongs to the technical field of animal biology. The building methodincludes: on the basis of porcine fetal fibroblast, utilizing CRISPR / Cas9 gene editing technology to knock out three genes GGTA1, CMAH and B4GalNT2 at the same time, and inserting three humanized modifying genes hCD55, hCD59 and hCD46 in a fixed point manner; combining somatic cell nuclear transfer technology to build an xenotransplantation donor base pig modified by the above six genes; on the basis of the base pig, knocking out gene B2M, and inserting three modifying genes hCD47, A20 and CTLA4-Ig in a fixed point manner at the same time; utilizing the somatic cell nuclear transfer technologyto build a islet cell and skin xenotransplantation donor pig modified by multiple genes; performing functional verifying on the donor pig to obtain the donor pig suitable for islet cell and skin xenotransplantation. A brand-new research concept and method are provided for developing the donor pig suitable for xenotransplantation of the islet cells and the skin, and clinical conversion and application of pigs serving in heterogeneous islet cell and skin transplantation are promoted.

Description

technical field [0001] The invention belongs to the field of animal biotechnology, in particular to a construction method suitable for islet cells and skin tissue xenotransplantation donor pigs. Background technique [0002] Islet cell transplantation is an effective and important approach for diabetic patients with complete pancreatic failure. At present, the isolation technology of pancreatic islets is relatively mature, but the problems of immune rejection and transplantation dysfunction have not been well solved, which has led to the lack of widespread application of this technology. [0003] When it comes to xenotransplantation, because pigs are easy to raise and breed, their organ sizes match those of humans, and their physiology and metabolism are very similar to those of humans, the possibility of comorbidities between humans and pigs is low, and there are few ethical and moral disputes involved. genetic modification to enhance the matching of donor tissues and orga...

AI Technical Summary

Problems solved by technology

[0004] Although pigs have been recognized as a source of human organ transplantation, xenogeneic tissue transplantation still faces the risk of cross-species disease infection, immunological incompatibility and functional impairment, making pig islet cells and skin tissue as xenografts Faced with severe challenges, it is not enough to transform into clinical application of medicine, making xenotransplantation stagnant for a long time

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