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Application of histone H3K79 methyltransferase inhibitor in improving development efficiency of animal round sperm injection embryos

A technology of methyltransferase and H3K79, which is applied in the field of medicine to achieve the effect of improving the development rate of blastocysts and improving the feasibility

Active Publication Date: 2021-09-17
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on the methylation modification function of H3K79 site mostly focuses on cell differentiation, DNA damage repair, telomere silencing, etc., and its role in improving the development efficiency of round sperm injection embryos in animals has not been reported yet.

Method used

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  • Application of histone H3K79 methyltransferase inhibitor in improving development efficiency of animal round sperm injection embryos
  • Application of histone H3K79 methyltransferase inhibitor in improving development efficiency of animal round sperm injection embryos
  • Application of histone H3K79 methyltransferase inhibitor in improving development efficiency of animal round sperm injection embryos

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1 Effect of Histone H3K79 Methyltransferase Inhibitors on Improving the Developmental Efficiency of Round Sperm Injection Embryos

[0107] 1. Oocyte Collection

[0108](1) Prepare egg storage dishes in advance: use imported Eppendorf embryo culture dishes with a diameter of 3.5cm, make eight 30μL culture drops with M16 medium for each dish, add 2.5mL paraffin oil, submerge the culture medium, and put them in the incubator (37°C, 5% CO 2 ) Pre-balanced for 3h.

[0109] (2) Select 10-week-old B6D2F1 female mice in the estrous period (F1 offspring obtained by mating DBA / 2N male mice with C57BL / 6N female mice, C57BL / 6N, DBA / 2N were purchased from Victoria River), body weight About 18~25g. Calculate the unit of hormone injection according to body weight, inject 8U per 20g of body weight; inject pregnant horse serum gonadotropin (PMSG) intraperitoneally at around 7:30 in the evening, and inject human chorionic gonadotropin 48 hours after PMSG starts (Humanchorioni...

Embodiment 2

[0132] Example 2 Effects of the concentration and action time of histone H3K79 methyltransferase inhibitors on the blastocyst development rate of embryos injected with round sperm

[0133] 1. The effect of the concentration of histone H3K79 methyltransferase inhibitor on the blastocyst development rate of round sperm injection embryos

[0134] This test process is identical with step 1, 2, and 3 in embodiment 1, and the difference is only in:

[0135] (1) The concentrations of the histone H3K79 methyltransferase inhibitor SGC0946 in the third step of the test process in the activation solution and the KSOM culture solution were 500 nM, 1 μM and 2 μM, respectively, and each treatment was repeated 3 times.

[0136] The result is as figure 2 Shown: the histone H3K79 methyltransferase inhibitor SGC0946 at a concentration of 1 μM is more effective in promoting the blastocyst development rate of round sperm injection embryos than the histone H3K79 methyltransferase inhibitor SGC09...

Embodiment 3

[0141] Example 3 Effect of Histone H3K79 Methyltransferase Inhibitors on Improving the Blastocyst Development Rate of Round Sperm Injection Embryos

[0142] This test process is identical with step 1, 2, and 3 in embodiment 1, and the difference is only in:

[0143] (1) The types and concentrations of DNA methyltransferases in the chemical activation solution containing histone H3K79 methyltransferase inhibitors and KSOM culture medium were different during the test: Figure 4 Middle: "ROSI" means without any treatment; "+DMSO" means adding the same amount of DMSO; "SGC0946-1μM" means SGC0946 with a concentration of 1 μM; "EPZ0047777-1 μM" means EPZ0047777 with a concentration of 1 77-5); "EPZ5676-1 μM" means EPZ5676 (CAS No.: 1380288-87-8) at a concentration of 1 μM.

[0144] The result is as Figure 4 Shown: Histone H3K79 methyltransferase inhibitor SGC0946 can significantly improve the blastocyst development rate of round sperm injection embryos.

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Abstract

The invention belongs to the field of medicine, and discloses application of a histone H3K79 methyltransferase inhibitor in improving the development efficiency of animal round sperm injection embryos. The invention discloses the application of the histone H3K79 methyltransferase inhibitor in improving the development efficiency of the animal round sperm injection embryos for the first time. Compared with an existing round sperm injection technology, the application has the advantages that under the condition that the transgenic risk is not introduced, the blastocyst development rate of the round sperm injection embryos can be remarkably increased through the histone H3K79 methyltransferase inhibitor, a good round sperm injection embryo in-vitro culture system is established, and the feasibility of the round sperm injection technology is improved.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to the application of a histone H3K79 methyltransferase inhibitor in improving the developmental efficiency of animal circular sperm injection embryos. Background technique [0002] There are no mature sperm or elongated sperm cells in the testes and epididymis of patients with severe azoospermia, so they cannot obtain genetic offspring by intracytoplasmic sperm injection (ICSI). The round spermatid injection (ROSI) technology is to inject round sperm into oocytes and fertilize oocytes through auxiliary activation, thereby helping these azoospermic patients obtain genetic offspring. Although the ROSI technique has been applied in a variety of animals including mice, rabbits, monkeys, and humans, there are still problems such as low implantation rate, high abortion rate, and low live birth rate. Therefore, improving the developmental potential of ROSI embryos can strongly promote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N5/075C12N5/076
CPCC12N5/0609C12N5/061C12N5/0604C12N2500/30
Inventor 赵小阳王京任少芳周偲马琳子李朝晖罗芳常港
Owner SOUTHERN MEDICAL UNIVERSITY
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