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Embryo antioxidation related lncRNA and application thereof

A technology of anti-oxidation and anti-oxidation ability, applied in the field of lncRNA function research, can solve the problems of lncRNA reporting without embryo anti-oxidation and developmental ability, and achieve the effect of improving developmental potential

Active Publication Date: 2021-04-27
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In particular, the lncRNA related to the antioxidant and developmental ability of embryos can provide technical reference for improving the developmental potential of embryos in vitro, but there is no lncRNA related to the antioxidant and developmental ability of embryos so far.

Method used

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  • Embryo antioxidation related lncRNA and application thereof
  • Embryo antioxidation related lncRNA and application thereof
  • Embryo antioxidation related lncRNA and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Preparation of Bovine IVF Embryos

[0038] The bovine ovary was collected from a local slaughterhouse, and the follicular fluid of follicles with a diameter of 2-6 mm was extracted. Pick cumulus-oocyte complexes under a stereomicroscope at 38.5°C, 5% CO 2 and 100% humidity in CO 2 Cultivate in the incubator for 22-24 hours. Mature oocytes were co-incubated for 8 hours in fertilization drops and thawed bovine sperm incubator. Afterwards, use vortex shaking for 2-3 minutes to remove the peripheral cumulus cells to obtain fertilized eggs, and place the fertilized eggs in the early culture medium containing 3mM glutathione (GSH) (109.5mM NaCl, 26.2mM NaHCO 3 , 3.1mM KCl, 1.19mM KH 2 PO 3 , 0.8mM MgCl 2 ·6H 2 O, 1.5mM glucose, 5mM calcium galactose, 0.4mM sodium pyruvate, 2% essential amino acids, 1% non-essential amino acids, 1mM L-glutamine, 6mg / mLBSA). After 48h, the embryos were transferred into the late culture medium containing 3mM GSH (109.5mM NaCl, 26.2mM NaH...

Embodiment 2

[0067] After overexpression of PANX1 in embryos, the antioxidant capacity of embryos was detected

[0068] Method for Overexpression of PANX1 Gene in Embryos

[0069] The overexpression plasmid of PANX1 gene was purchased from Shanghai Gemma Pharmaceutical Technology Co., Ltd. The vector was linearized and transcribed in vitro, and the obtained PANX1 mRNA (concentration 10 μM) was injected into 1-cell stage in vitro fertilized bovine embryos, each embryo was injected with 10 pL.

[0070] Detection method of GSH content in embryo:

[0071] The fluorescent dye Cell Tracker Blue CMF2HC (Life Technology) was used to stain 2-cell stage embryos, 4-cell stage embryos, 8-cell stage embryos, morula and blastocysts (including control group and overexpression group) to determine which of GSH content. Embryos were washed three times with 0.1% BSA-DPBS, placed in a staining solution with a concentration of 10 μM, incubated at 38.5°C in the dark for 15 minutes, then taken out, washed thr...

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Abstract

The invention provides embryo antioxidation related lncRNA and application thereof, and belongs to the technical field of lncRNA function research. A nucleotide sequence of the lncRNA CUFF.97956.1 is as shown in SEQ ID No.1; a target gene of the lncRNA CUFF.97956.1 is a PANX1 gene; the expression quantity of the lncRNA CUFF.97956.1 in the bovine embryo is remarkably reduced after glutathione treatment; the lncRNA CUFF.97956.1 and the target gene thereof are related to the antioxidant and developmental capacity of the embryo; and the lncRNA CUFF.97956.1 and the target gene thereof provide technical support for improving the developmental potential of the in-vitro embryo.

Description

technical field [0001] The invention belongs to the technical field of lncRNA function research, and in particular relates to an lncRNA related to embryo anti-oxidation and its application. Background technique [0002] As we all know, the development rate of mammalian in vitro embryos is far lower than that of in vivo embryos, and its low pregnancy rate and high fetal abortion rate have become restrictive factors for the widespread use of bovine in vitro embryos in the cattle industry, which in turn affects the income of farmers and herdsmen and my country's animal husbandry. healthy, rapid and sustainable development of the industrial economy. In order to improve the quality of embryos in vitro, the antioxidant glutathione (GSH) is often used to scavenge oxygen free radicals in embryos, which plays a vital role in embryonic development. [0003] Existing studies have shown that a large number of lncRNAs have been found in oocytes and embryos, and they play an important rol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/113C12N15/11
CPCC12Q1/6888C12Q2600/158C12Q2600/178
Inventor 杜卫华田雅晴
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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