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Vector and method for increasing cattle cloning efficiency based on histone methylation level modification

A methylase and carrier technology, applied in biochemical equipment and methods, other methods of inserting foreign genetic materials, botanical equipment and methods, etc. Growth rate and quality, effect of promoting stable translation

Pending Publication Date: 2018-08-17
NORTHWEST A & F UNIV
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Problems solved by technology

[0004] Although epigenetic modifications such as histone methylation are widely present in animal embryo development, no studies have revealed the relationship between epigenetic modifications and endogenous proteins from the perspective of differences in epigenetic modifications between cloned and fertilized embryos. The relationship between expression levels leads to the lack of clear and effective solutions for how to improve the efficiency of animal embryo development in vitro, especially the use of differential expression of KDM4 family genes to improve the developmental rate of bovine somatic cell clone embryos, the number of blastocyst cells and the birth rate Haven't seen the report yet

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  • Vector and method for increasing cattle cloning efficiency based on histone methylation level modification
  • Vector and method for increasing cattle cloning efficiency based on histone methylation level modification
  • Vector and method for increasing cattle cloning efficiency based on histone methylation level modification

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[0042]The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments. The examples are by way of illustration, not limitation, of the invention.

[0043] By immunofluorescence staining, bovine somatic cell clone embryos were found to have abnormally high levels of histone H3 lysine K9 3-methylation (H3K9me3) and 2-methylation (H3K9me2) at the early stage of zygotic activation (8-cell stage), and figure 1 The results showed that bovine somatic cell clone embryos had abnormally high levels of histone H3K9me3 and H3K9me2 epigenetic modifications at the 8-cell stage compared with in vitro fertilized embryos. In addition, by real-time fluorescent quantitative PCR, it was determined that the abnormally high level of histone methylation in bovine somatic cell clone embryos was related to its own abnormally low expression level of the KDM4E gene ( figure 2 The results showed that KDM4A, KDM4B, KDM4C and other genes in b...

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Abstract

The invention discloses a vector and method for increasing cattle cloning efficiency based on histone methylation level modification. The vector comprises the in-vitro transcription template vector containing a cattle KDM4E gene. A large amount of mRNA of the cattle KDM4E gene can be obtained by subjecting the vector to in-vitro transcription reaction, and the mRNA is injected to a successfully fused and activated bovine somatic cell cloning embryo to allow the embryo to overexpress cattle KDM4E protein. The method has the advantages that histone H3K9me3 and H3K9me2 epigenetic modification level of the somatic cell cloning embryo during zygotic activation can be effectively lowered, so that the development rate and quality of later bovine somatic cell cloning embryo can be increased evidently, the method can be used for efficiently producing the bovine somatic cell cloning embryo in vitro, and the birth rate of cloning cattle after embryo transplanting is increased evidently.

Description

technical field [0001] The invention belongs to the technical field of animal somatic cell cloning, and relates to a carrier and a method for improving bovine cloning efficiency based on the modification of histone methylation level. Background technique [0002] Somatic cell cloning is a technology with great research and commercial value, which can be applied to therapeutic cloning, disease models, human organ transplantation, animal transgenic research, protection of endangered animal breeds, expansion of excellent livestock breeds, etc. But so far the efficiency of somatic cell cloning is still not high, which significantly restricts the wide application of this technology. [0003] At present, it is believed that the main reason for the low cloning efficiency is that the nucleus of the donor somatic cell is not completely reprogrammed by the cytoplasm of the recipient ooplasm, that is, the reprogramming fails or is incomplete. This reprogramming process does not involv...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/89C12N15/877
CPCC12N9/0071C12N15/63C12N15/8771C12N15/89C12Y114/11027
Inventor 张涌刘鑫张景程高元鹏邢徐鹏周川
Owner NORTHWEST A & F UNIV
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