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Expression inhibitor of porcine HDAC2 gene and application of expression inhibitor

A gene expression and inhibitor technology, applied in the biological field, can solve the problems of restricting the popularization and application of somatic cell cloning pigs, and low production efficiency, and achieve the effect of increasing the number of cells in blastocysts, reducing negative effects, and improving developmental performance

Active Publication Date: 2020-04-03
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Pig somatic cell cloning technology has important application value in basic scientific research, animal breeding and medical research, but the overall production efficiency of somatic cell cloned pigs is still low, and the cloning efficiency of pigs (relative to the efficiency of the number of reconstructed eggs) No more than 0.3%, which greatly limits the further promotion and application of somatic cell cloned pigs

Method used

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  • Expression inhibitor of porcine HDAC2 gene and application of expression inhibitor
  • Expression inhibitor of porcine HDAC2 gene and application of expression inhibitor
  • Expression inhibitor of porcine HDAC2 gene and application of expression inhibitor

Examples

Experimental program
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Effect test

Embodiment 1

[0035] 1. Design and synthesis of porcine HDAC2 gene and siRNA of porcine HDAC2 gene

[0036] According to the porcine HDAC2 gene sequence (sequence number: XM_001925318.6), referring to the principle of siRNA design, three pairs of siRNA were designed and synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd. with the corresponding target sites of the porcine HDAC2 gene, respectively denoted as HDAC2-siRNA1, The sequences of HDAC2-siRNA2 and HDAC2-siRNA3 are shown in Table 1 below:

[0037] Table 1 siRNA sequence

[0038]

[0039] 2. The inhibitory effect of siRNA on genes

[0040] 2.1 siRNA solution preparation

[0041] Centrifuge the tubes containing the dry siRNA powder, and then add 150 μL of sterile, DNase & RNase-free embryo injection-grade water to each tube (purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.). Concentration of 20mM, cap the bottle tightly and shake to dissolve. After centrifugation, dispense 5 μL / tube into sterile, DNase & RNas...

Embodiment 2

[0064] 1. The effect of CAY10683 on the developmental performance of pig somatic cell cloned embryos

[0065]The small molecule CAY10683, which specifically inhibits HDAC2, was added to the culture medium to incubate and culture the reconstituted embryos of pig clones at the 1cell stage for 24 hours, and then replaced with normal medium to continue the culture. The NC group was supplemented with the same amount of untreated normal medium. The cleavage rate of cloned and reconstituted embryos in the experimental group was observed on day 2 and the blastocyst development rate on day 7, and nuclear fluorescent staining was performed on blastocyst cells, and the total number of cells was counted. The results are shown in Table 3.

[0066] Table 3 Effects of different concentrations of CAY10683 on the developmental performance of pig somatic cell cloned embryos

[0067]

[0068] As can be seen from the results in Table 3, compared with the NC group, the CAY10683 treatment grou...

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Abstract

The invention discloses an expression inhibitor of a porcine HDAC2 gene, which can effectively improve the development rate of a porcine cloned and reconstructed embryo and the embryo quality in a blastocyst period, which can effectively improve the development performance of the porcine cloned and reconstructed embryo, so that the cloning efficiency of porcine somatic cell cloning can be improved. The expression inhibitor of the porcine HDAC2 gene can be selected from at least one of siRNA for inhibiting the expression of the porcine HDAC2 gene and a compound capable of inhibiting the expression of the porcine HDAC2 gene.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an expression inhibitor of porcine HDAC2 gene and its application in preparing a product capable of improving the developmental performance of porcine clone reconstituted embryos. Background technique [0002] Somatic Cell Nuclear Transfer (SCNT), also known as somatic cell cloning technology, is a commonly used technology in animal cell engineering technology. It uses somatic cells as nuclear donors and uses special artificial means (micromanipulation, electrofusion, etc.) etc.) Transplant somatic cell nuclei into enucleated oocytes to generate reconstituted embryos. After in vitro fusion activation and culture to a certain stage, the reconstituted embryos are then transplanted into surrogate mothers so that they develop into the same genetic material as the donor cells. technology of cloning offspring. Animals obtained by somatic cell nuclear transfer become cloned ani...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/877
CPCC12N15/113C12N15/8778C12N2310/14
Inventor 吴珍芳蔡更元王兴旺李紫聪吴宵招华兴杨柳松
Owner SOUTH CHINA AGRI UNIV