Method for application of H3K27me3 and demethylase KDM6A/B thereof to nuclear transfer reconstructed embryo of mouse

Abstract

The invention discloses a method for application of H3K27me3 and demethylase KDM6A / B thereof to a nuclear transfer reconstructed embryo of a mouse. With the application method, the difference between an H3K27 trimethylation mode in a parthenogenetic embryo in each period before implantation and that in a clonal embryo, and the influence of the difference in the mode on the developmental capacity of the clonal embryo are analyzed, and the method verifies that fine adjustment of KDM6A and KDM6B can improve the blastocyst development rate of the cloned embryo and the birth rate of a cloned animal.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an application method of H3K27me3 and its demethylase KDM6A / B in mouse nuclear transfer reconstituted embryos. Background technique [0002] Somatic cell nuclear transfer refers to the technique of injecting somatic cell nuclei into the enucleated oocyte cytoplasm to obtain reconstructed embryos, which eventually develop into offspring; cell reprogramming can be achieved through nuclear transfer, cell fusion, specific transcription factors and culture. However, the only reprogramming method that can induce cells to regenerate into individuals is somatic cell nuclear transfer. Other methods can only induce induction at the cellular, molecular or biochemical level, and they all have some shortcomings. Since the establishment of somatic cell nuclear transfer technology, cloning efficiency has been at a low level, only a few reconstituted embryos can develop to term birth, usually only 1...

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Problems solved by technology

[0008] It has been reported that in mouse embryonic development and cell differentiation, KDM6A has an important regulatory effect on key genes affecting embryonic development; like KDM6A, KDM6B is also an important demethylase of H3K27me3, encoded by autosomes, in Studies on mice have shown that KDM6B is involved in the transformation of white and brown fat by regulating the activity of key fat genes; after bone injury, KDM6B can regulate the differentiation of osteoclasts and may limit the pathogenesis of osteoporosis, and is associated with There is a certain relationship between skin repair; when nerve cells are damaged, KDM6B can limit the excessive proliferation and regeneration of Schwann cells, thereby preventing the generation of neurofibromas. In addition, KDM6B also plays a regulatory role in pro-inflammatory and anti-inflammatory responses. In summary, the demethylase activity of KDM6A and KDM6B is very important in cell differentiation, not only involved in cell cycle regulation, macrophage differentiation, neural stem cell differentiation, but also can regulate cell reprogramming process, participate in embryonic stem cell differentiation , regulating the expression of genes related to embryonic development and other aspects, H3K27me3 histone demethylase KDM6A / B was discovered in 2007, and related research mainly focused on the biochemical mechanism of demethylation, studying KDM6A / B Its role in the development of nuclear transfer reconstituted embryos has not been reported

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