Breeding method and application of transgenic pig

Abstract

The invention discloses a method for breeding transgenic pigs. The breeding method of the transgenic pig at least includes introducing the primary somatic cells of the pig into the cell reprogramming switch gene and / or miR302 / 367 cluster encoded by messenger RNA, so that it can be induced into RNA-induced pluripotent stem cells; Cas9-gRNA and CRE simultaneously Under the action of the original recombinase, several human gene fragments are knocked in while knocking out the toxic gene of the pig, and the human vWF gene is used to replace the endogenous vWF gene of the pig in situ, and the PERV virus is inactivated, and there is no immune rejection and blood coagulation incompatibility genes; screened out clean on-target cell lines without off-target effects and cultured them into pig somatic cells; processed pig somatic cells with somatic cell nuclear transfer technology to obtain transgenic pigs. The transgenic pig obtained by the invention has the advantages of permanent inability to express the PERV virus, no immune rejection to the human body, compatibility with blood coagulation, etc., and can be used as a donor of human heterologous organs, tissues and cells.

Description

technical field [0001] The invention relates to the field of transgenic technology, in particular to a breeding method and application of a transgenic pig. Background technique [0002] Organ transplantation is an effective treatment for human terminal organ failure, but it is severely limited by the shortage of donor organs, and a large number of people die each year while waiting for organs. In addition, the high cost of organ transplantation also limits the number of human organ transplant operations. In terms of the number of donor organs and the cost of organ transplantation, people think of heterologous transplantation, that is, transplanting the corresponding organs of other animals into the human body that needs organ transplantation. Among many animal organs, the size and physiological functions of pig organs are similar to those of humans, so they are recognized as the best donors for xenotransplantation. However, there is porcine endogenous retrovirus (PERV) in ...

AI Technical Summary

Problems solved by technology

[0008] The present invention aims at the existing problems of PERV virus expression, human immune rejection and coagulation incompatibility in the existing human-pig allografts, as well as the cumbersome screening process, low cloning rate, incomplete genetic modification and inability to form pig iPS in the existing solutions to the above problems Problems such as embryos, providing a breeding method for transgenic pigs

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