Method for obtaining cloned pigs with modified genes through continuous cloning

Abstract

The invention relates to a method for obtaining cloned pigs with modified genes through continuous cloning and belongs to the field of animal reproductive biology.According to the method, tissue fibroblasts of obtained genetically modified pigs serve as donors, donor cells are transferred into enucleated porcine oocytes through the somatic cell nuclear transfer to build a genetically modified pig reconstructed embryos, then electro-fusion and electro-activation are conducted, the donor cells are transferred into estrous surrogate sow fallopian tube expanded portions after being cultured in a pig embryo in-vitro culture solution for 3 h, and after gestation of surrogate sows last for 114 d, a large scale of genetically modified pig individuals are obtained through delivery.The method can solve the problems that in the process that genetically modified individuals are obtained through the gene editing technology, target genetically modified individuals are low in efficiency, and difference between the target genetically modified individuals is large, so that genetically modified cloned pig individuals stable in target gene expression and phenotype can be efficiently produced on a large scale.

Description

technical field [0001] The invention relates to a method for producing genetically modified cloned pigs through continuous cloning technology, belonging to the field of reproductive biology. Background technique [0002] In recent years, the in-depth research and construction of genetically modified pigs as human disease models has not yet been comprehensive, and there are shortcomings such as low construction efficiency and narrow application range. Animal disease models established from the perspective of animal genetically modified cloned pigs are conducive to in-depth exploration of human-related diseases It has important reference value for the construction of clinical models of human diseases and clinical transformation applications, especially for the study of genetically modified cloned pigs obtained by serial cloning methods as animal disease models. The repeatability of the source, the reliability of the research and the stability of the genetics, the pathological ...

AI Technical Summary

Problems solved by technology

[0002] In recent years, the in-depth research and construction of genetically modified pigs as human disease models has not yet been comprehensive, and there are shortcomings such as low construction efficiency and narrow application range. Animal disease models established from the perspective of animal genetically modified cloned pigs are conducive to in-depth exploration of human-related diseases It has important reference value for the construction of clinical models of human diseases and clinical transformation applications, especially for the study of genetically modified cloned pigs obtained by serial cloning methods as animal disease models. The repeatability of the source, the reliability of the research and the stability of the genetics, the pathological environment, physiological conditions, biochemical mechanism and the mechanism of the occurrence and development of the disease in pigs are very close to those of humans. The genetically modified clones established by this method The pig model provides an important model reference for the study of human-related diseases

[0003] At present, although the pig gene targeting technology can change the specific gene expression pattern of animals, it is extremely difficult to operate. Although the pig transgenic technology is simple to operate and the cycle is short, the efficiency of obtaining individual pigs modified with the target gene is low, and there are large differences among individuals. , it is difficult to achieve the desired effect

Images