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Method for conducting PGS and methylation analysis on biopsy cells by means of DedscRRBS analysis method

A technology of methylation and analysis method, which is applied in the fields of biomedicine and molecular cell biology, can solve the problems of large SD value, neglect of epigenetic changes, and small amount of effective data, so as to achieve the effect of improving the success rate

Pending Publication Date: 2019-06-28
北京中科遗传与生殖医学研究院有限责任公司
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Problems solved by technology

[0009] (1) Traditional PGS only evaluates from the chromosomal state, ignoring the very important epigenetic changes in the early embryo, and cannot provide more references for a more comprehensive evaluation of the embryonic developmental potential;
[0010] (2) Both single-cell whole-genome bisulfite sequencing (scWGBS) and simplified expression bisulfite sequencing (scRRBS) based on single-enzyme digestion (Msp I) processing have a relatively low genome matching rate, and the obtained effective data The amount is relatively small; and when performing PGS analysis, the SD value is large, and a clear result of chromosomal aneuploidy cannot be obtained

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  • Method for conducting PGS and methylation analysis on biopsy cells by means of DedscRRBS analysis method
  • Method for conducting PGS and methylation analysis on biopsy cells by means of DedscRRBS analysis method
  • Method for conducting PGS and methylation analysis on biopsy cells by means of DedscRRBS analysis method

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Embodiment Construction

[0038] In order to further understand the content of the present invention, the present invention will be described in detail below in conjunction with specific embodiments.

[0039] The experimental methods that do not indicate specific conditions in the following examples are usually according to conventional conditions, or according to the conditions suggested by the manufacturer.

[0040] Two cases of blastocyst biopsy cells (sample 1 and sample 2) were taken for DedscRRBS-PGS sequencing analysis, and the specific operations were as follows:

[0041] 1. Acquisition of blastocyst biopsy cells: Obtain fertilized eggs by intracytoplasmic sperm injection (ICSI), culture the fertilized eggs in blastomere medium (G1 medium) for 3 days to the blastomere stage, and then Embryos were transferred to blastocyst medium (G2 medium) and cultured until the 5th day until they developed to the stage of blastocyst maturity. Take 3-5 trophectoderm cells away from the inner cell mass of blas...

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Abstract

The invention relates to a method for parsing the blastocyst ectoderm cell DNA methylation status by means of a phenogenetics analysis technology by taking 'test tube baby' blastocyst ectoderm cells as raw materials and adopting a double-enzyme digestion single-cell reduced-representation bifulfite sequencing technology and conducting preimplantation genetic screening on the chromosome state. Thedevelopment potential of an embryo is comprehensively evaluated from various angles, a new reference is provided for selecting the 'right' embryo in assisted reproduction, and strong support is provided for increasing the success rate of the 'test-tube baby' cycle.

Description

technical field [0001] The invention relates to the fields of biomedicine and molecular cell biology, in particular to a method for performing single-cell double-enzyme digestion and simplified representative bisulfite sequencing on trophoblast biopsy samples of test-tube baby embryos, and using epigenetic analysis technology to analyze cysts DNA methylation status of ectoderm cells, and preimplantation genetic screening for chromosomal status. Background technique [0002] As a medical invention with cross-age significance to the history of human reproduction, test-tube baby technology is considered to be a "milestone in the development of modern medicine". Since its birth in 1978, this technology has brought hope to countless families with fertility disorders all over the world in the past 40 years, and has also found effective solutions to the increasingly prominent human fertility problems. With the rapid development of technology, test-tube babies have also experienced...

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Application Information

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IPC IPC(8): C12Q1/6858
Inventor 张癸荣费嘉金治平王云云蔡旺庭刘威达
Owner 北京中科遗传与生殖医学研究院有限责任公司
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