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A method for improving mammalian cloning efficiency based on obtaining specific donor cells

A donor cell, efficient technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve the problems of low birth efficiency of mammalian cloned embryos

Active Publication Date: 2020-12-08
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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  • A method for improving mammalian cloning efficiency based on obtaining specific donor cells
  • A method for improving mammalian cloning efficiency based on obtaining specific donor cells
  • A method for improving mammalian cloning efficiency based on obtaining specific donor cells

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Embodiment Construction

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

[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 into a 37°C constant temperature water bath, and shake quickly until the cell cryopreservation solution is completely dissolved. Carefully wipe the cryopreservation tube 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 10cm cell culture plate, then add 8 mL of 10% FBS cell culture medium, shake the culture plate gently to spread the cells evenly, and place them in a 37°C, 5% CO2 cell incubator for culture. After the cells adhered to...

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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 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 science, but the efficiency of this technology is still very low. This severely limits the application of this technique. Taking pig cloning technology as an example, the low efficiency of this technology is mainly reflected in the fact that the development and birth efficiency of pig cloned embryos is only about 0.5% (number of cloned pigs born / number of cleavage stage cloned embryos transferred to surrogate sows), which is much lower than that of pig cloned embryos. The developmental birth rate of embryos fertilized in vivo, which is about 75%. Studies have shown that one of the main reasons w...

Claims

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Application Information

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Patent Type & Authority Patents(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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