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
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Examples
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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Description
Claims
Application Information
- IPC
- A61K48/00; A61K31/7088; A61K38/17; A61K39/395; A61P15/18; A61P15/08
- Inventors
- 王健; 申庆祥