Method for improving mammal cloning efficiency based on acquisition of specific donor cells

A donor cell, mammalian technology, applied in the fields of botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve the problem of low birth efficiency of mammalian cloned embryo development and so on

Active Publication Date: 2018-10-30
SOUTH CHINA AGRI UNIV
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Problems solved by technology

Studies have shown that the DNA methylation level of XIST-DMR in mammalian cloned embryos at the blastocyst stage is significantly higher than that of in vivo fertilized embryos, and this may be one of the reasons why the development and birth efficiency of mammalian cloned embryos is lower than that of in vivo fertilized embryos main reason

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  • Method for improving mammal cloning efficiency based on acquisition of specific donor cells
  • Method for improving mammal cloning efficiency based on acquisition of specific donor cells
  • Method for improving mammal cloning efficiency based on acquisition of specific donor cells

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Embodiment Construction

[0021] The present invention will be further described in detail below in conjunction with specific embodiments.

[0022] The XIST-DMR fragment sequence involved in the present invention is derived from GenBank: KC149530.1.

[0023] 1 Donor cell clone group culture

[0024] 1.1 Cell recovery

[0025] Place the cryopreservation tube containing Duroc boar 510D cells in a 37°C constant temperature water bath and shake it quickly until the cell cryopreservation solution is completely dissolved. Wipe the cryopreservation tube carefully with alcohol cotton, transfer the cell suspension to a 15 mL centrifuge tube, centrifuge at 800 rpm for 5 min, remove the supernatant, and add 2 mL of 10% FBS cell culture medium to resuspend the cells. Transfer the cell suspension to a 10 cm cell culture plate, and then add 8 mL of 10% FBS cell culture medium, gently shake the culture plate to spread the cells, and place the cells in a 37°C, 5% CO2 cell incubator for culture. After the cells adhere to the...

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Abstract

The invention discloses a method for improving mammal cloning efficiency based on acquisition of specific donor cells. According to the method, the donor cells with low-level DNA (deoxyribonucleic acid) methylation in XIST-DMR (X-inactive specific transcript-deuteron magnetic resonance) are acquired and used for nuclear transfer experiments, so that the mammal cloning efficiency is improved. Cloned embryos generated by the method for improving the mammal cloning efficiency based on acquisition of the specific donor cells are lower in XIST-DMR DNA methylation level at a blastula stage, so thatthe development birth rate of the cloned embryos is remarkably increased.

Description

Technical field [0001] The present invention relates to the field of animal biotechnology, in particular to a method for improving mammalian cloning efficiency based on obtaining specific donor cells. Background technique [0002] Mammalian somatic cell nuclear transfer (cloning) technology has important application value in the fields of animal husbandry, human medicine, pharmacy and life sciences, but the efficiency of this technology is still very low. This severely limits the application of this technology. Take the pig cloning technology as an example. The low efficiency of this technology is mainly reflected in the development and birth efficiency of pig cloned embryos, which is only about 0.5% (the number of cloned pigs born / the number of cloned embryos transferred to the surrogate sow), which is much lower than that of pigs. The developmental birth efficiency of in vivo fertilized embryos is about 75%. Studies have shown that one of the main reasons that the development...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/873A01K67/027
CPCA01K67/0271A01K2227/108C12N15/873
Inventor 吴珍芳李紫聪吴霄赵成成王兴旺
Owner SOUTH CHINA AGRI UNIV
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