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Function of E4BP4 gene inplantation course and use thereof

A technology of E4BP4 and gene, which is applied to the function and application field of E4BP4 gene in the process of embryo implantation, and can solve the problems of undiscovered research reports and so on

Inactive Publication Date: 2007-08-29
SHANGHAI INST OF PLANNED PARENTHOOD RES
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] Although it has been reported that E4BP4 gene is also expressed in rat embryos, and the expression level is the highest in embryonic neurons, and it is an embryonic endogenous regulatory factor involved in the regulation of motor neuron survival and growth (Junghans et al. , D., et al., 2004, The CES-2-related transcription factor E4BP4 is an intrinsic regulator of motoneuron growth and survival. Development, 131: 4425-4434.), but so far no information about the E4BP4 gene and its expression products involved in regulation Research Report on Embryo Implantation Process

Method used

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  • Function of E4BP4 gene inplantation course and use thereof
  • Function of E4BP4 gene inplantation course and use thereof
  • Function of E4BP4 gene inplantation course and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0062] Expression of E4BP4 gene

[0063] The present inventors detected and compared the expression status of the E4BP4 gene in periimplantation uterine tissues of mice from the levels of transcription and translation. Adult ICR mice aged 8-10 weeks were taken, and the female and male mice were housed in a cage at a ratio of 2:1. The vaginal suppository of the female mice was checked the next morning to determine whether they had mated, and the day when the suppository was seen was regarded as the first day of pregnancy. sky. For the non-pregnant female mice, as well as the mice on the 1st day to the 4th day of pregnancy, they were sacrificed by neck dislocation at 8:00 am on the same day, and their uterine tissues were collected. For the mice on the 5th day of pregnancy and the 6th day of pregnancy, the mice were injected with Chicago blue solution (1% normal saline, 0.1ml per mouse) from the tail vein at 8:00 in the morning of the same day, and then killed by dislocation of...

Embodiment 2

[0068] The antisense oligonucleotide chain of E4BP4 mRNA inhibits embryo implantation in mice

[0069]For the 20 nucleotide sequence in the mouse mRNA coding region: 5'-AUCAAAAAGGAGCCCGCACC-3' (SEQ ID NO: 5), the inventors synthesized its antisense DNA fragment: 5'-GGTGCGGGCTCCTTTTTGAT-3' (SEQ ID NO: 5) NO: 6). At the same time, a random sequence control DNA fragment: 5'-TGAGGAGCAGCCCACAGAAG-3' (SEQ ID NO: 7) was synthesized. Take the Mirus TransIT of Madison Company of the United States R Polymer is used as a transfection agent, and the in vivo E4BP4 gene function blocking experiment was carried out, the method is as follows:

[0070] (1) Add 10μL (1μg / μL) Mirus TransIT R After mixing Polymer solution and 20 μL Delivery solution, add 10 μL antisense DNA solution, 10 μL control DNA solution, and 10 μL DNA-solution respectively to prepare three different injection solutions;

[0071] (2) From 3:00 to 4:00 p.m. on the fourth day of pregnancy, inject the injection solution ...

Embodiment 3

[0079] Example 3 Screening of antagonists of E4BP4

[0080] According to the method described in Example 2, the following two groups of substances (replacing antisense nucleic acid and transfection agent) were administered to pregnant mice, and then the in vivo inhibitory effect on blastocyst implantation was observed: (a) candidate substance+E4BP4 factor; (b) E4BP4 factor.

[0081] If the blastocyst implantation rate of group (a) is statistically significantly lower than that of group (b), it indicates that the candidate substance is an antagonist of E4BP4.

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Abstract

The present invention discloses a new application of E4BP4 gene, it can be used as regulator for regulating embryo nidation (embryo implantation). In prophase of embryo nidation it can block expression of E4BP4 gene or close the biological function of its expression product E4BP4 protein, so that it can inhibit embryo nidation, otherwise it can promote nidation; in anaphase of embryo nidation it can block expression of E4BP4 gene or close the biological function of its expression product E4BP4 protein, so that it can promote embryo nidation, otherwise, it can inhibit nidation.

Description

technical field [0001] The invention belongs to the field of biotechnology and medicine, in particular, the invention relates to the function and application of the E4BP4 gene in the embryo implantation process. Background technique [0002] Embryo implantation (also known as embryo implantation) is an important step unique to the reproductive process of mammals. It refers to the interaction between the embryo in the activated state and the uterus in the accepting state, and finally leads to the establishment of tightness between the embryonic trophoblast and the endometrium. contact process. It is a reproductive physiological step unique to mammals and humans, and it is also an ideal link for fertility regulation. The implantation process is a complex physiological process involving an extremely complex and delicate synergy between the embryo and the uterus, including the development and differentiation of the embryo, the synchronous changes of the endometrium, and the int...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/7088A61K38/17A61K39/395A61P15/18A61P15/08
Inventor 王健申庆祥黄哲平
Owner SHANGHAI INST OF PLANNED PARENTHOOD RES
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