Cloned embryo treatment liquid, use method and uses thereof

A technology for cloning embryos and treatment solution, which is applied in the field of treatment solution for cloning embryos, and can solve problems such as low efficiency

Active Publication Date: 2017-10-03
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because of this advantage, the use of ESc for somatic cell cloning often has a high cloning efficiency, but its efficiency is still not high

Method used

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  • Cloned embryo treatment liquid, use method and uses thereof
  • Cloned embryo treatment liquid, use method and uses thereof
  • Cloned embryo treatment liquid, use method and uses thereof

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Embodiment Construction

[0029] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0030] Experimental Materials

[0031] 1. Cell lines and pig cloned embryos

[0032] Pig adult fibroblasts (513D) were collected from breeding pigs provided by Guangdong Wen's Breeding Pig Technology Co., Ltd. Shuitai Original Pig Farm; pig cloned embryos were prepared in the cloning laboratory of Guangdong Wen's Breeding Pig Technology Co., Ltd.

[0033] 2. Main reagents and consumables

[0034]RNA extraction kit E.Z.N.A.Total RNA Kit I was purchased from OMEGA Company; embryo RNA extraction kit Qiagen RNeasy Plus Micro Kit (50) was purchased from Qiagen Company; reverse transcription kit PrimeScriptTMRTreagent Kit with gDNA Eraser was purchased from TaKaRa Company; Quantitative PCR kit SYBR SelectMaster Mix was purchased from Life Technologies; Taq enzyme for common PCR was purchased from Guangzhou Dongsheng Biotechnology Company; DL500DNA Marker was purc...

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Abstract

The present invention discloses a cloned embryo treatment liquid, which comprises BIX-01294 and a PZM culture liquid, wherein the BIX-01294 concentration in the PZM culture liquid is 5-500 nM, and 500 mL of the PZM culture liquid comprise 3.156 of NaCl, 1.053 g of NaHCO3, 0.373 g of KCl, 0.024 g of KH2SO4, 0.049 g of MgSO4, 0.308 g of calcium lactate, 0.011 g of pyruvic acid, 0.073 g of L-glutamic acid, 0.273 g of hypotaurine, 10 mL of BME, 5 mL of MEM, 0.033 g of penicillin, 0.025 g of streptomycin, and the balance of Sigma water. According to the present invention, with the application of the cloned embryo treatment liquid using the technical scheme to perform the culture treatment on the cloned embryo, the gene expression of SUV39H1, SETdb1, DNMT1 or DNMT3a in the cell can be significantly reduced, and the H3K9me2 levels of 2cell and 4cell at the early embryo development stage can be significantly down-regulated.

Description

technical field [0001] The invention relates to somatic cell cloning technology, in particular to a treatment solution for cloning embryos, a method for using the treatment solution and the use of the treatment solution. Background technique [0002] Somatic cell nuclear transfer (SCNT), also known as somatic cell cloning, is an assisted reproductive technology used to produce animals. SCNT refers to the transplantation of donor somatic cells into the cytoplasm of enucleated oocytes, through fusion and activation to obtain reconstructed embryos, allowing the cells to resume division and continue to develop to obtain offspring with the same genotype as the donor cells. [0003] Through somatic cell cloning technology, the nucleus of a single differentiated somatic cell can be cultured in an in vitro environment to obtain a whole biological organism, which has great potential for agricultural technology, biomedicine industry, and protection of endangered species (Yang et al., ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N5/075
CPCC12N5/0604C12N5/0609C12N2500/05C12N2500/30C12N2500/32C12N2500/33
Inventor 吴珍芳杨林雪杨洋赵成法贺晓燕蔡更元李紫聪刘德武
Owner SOUTH CHINA AGRI UNIV
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