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Novel application of gene TBX3

A new use, gene technology, applied in the direction of peptide source, peptide, animal/human peptide, etc., can solve the problem of unclarified TBX3 expression change regulation trophoblast cells, etc., to improve the efficiency of embryo implantation and the success rate of assisted reproduction, expand The effect of testing standards

Pending Publication Date: 2020-01-10
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, previous findings have not clarified the changes in TBX3 expression during the periimplantation period and the regulation of trophoblast cell function by TBX3.

Method used

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  • Novel application of gene TBX3
  • Novel application of gene TBX3
  • Novel application of gene TBX3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Construct a new model SCBAV based on the informatics model to detect the development of trophoblast cells in the peri-implantation period. Total variation of at least 40% of the number); 2. Utilize Gap statistics and K-means clustering analysis of cell subgroups at each time point, wherein Gap statistics determine the number of cells in each subgroup at a specific time point, K-means Clustering classifies the number of clusters determined by Gap statistics; 3. Constructs cell lineage trajectories by iteratively associating the most correlated subpopulations between two time points.

[0017] Further, to detect genes that conform to the model in the peri-implantation trophoblast single-cell transcriptome, the gene needs to conform to two parameters of the model: 1. The regulatory factor must have a stable expression difference (expression) in the two lineages whose trajectories diverge ; 2. High degree of expression difference (dispersion) in the cell population before di...

Embodiment 2

[0022] The method and model of in vitro knockdown of TBX3 gene and control trophoblast cells JEG-3 in differentiating into syncytiotrophoblast cells

[0023] In vitro 8-bromo-cyclic AMP (8-Br-cAMP) induced differentiation of trophoblast cell line - choriocarcinoma cell line JEG-3 into syncytiotrophoblast cells, including detecting the number of syncytia formation and syncytial cell markers Expression of hCGβ.

[0024] Further, the method for knocking down the expression of TBX3 is a lentivirus targeting control sequence and TBX3 sequence, and the target sequence is as follows:

[0025]

[0026] The specific method is as follows: construct a lentiviral plasmid containing the target sequence, transfect it into 293T cells, discard the culture medium after mixing and culturing for 8 hours, add PBS buffer to wash, replace with fresh medium, continue to cultivate, and collect Virus culture solution, centrifuged to obtain lentivirus containing target control sequence and TBX3 seq...

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Abstract

The invention discloses a novel application of a gene TBX3. The novel application is characterized in that the gene has application in regulating trophoblastic cells to be differentiated into plasmoditrophoblast cells. The application has the advantages that a trophoblastic cell differentiation means can be supplemented, quality testing standards for trophoblastic cells and placentas are expandedand embryo implantation efficiency and an assisted reproduction success rate are improved.

Description

technical field [0001] The present invention relates to a new application of gene TBX3. Background technique [0002] The pre-implantation development of mammalian embryos mainly goes through several important cleavage and differentiation processes, including the first cleavage of fertilized eggs, the establishment of cell polarity and compaction of morula, and the differentiation of blastocysts into different cell lineages. Late blastocysts consist of three cell lineages: trophectoderm, primitive endoderm, and ectoderm. The endoderm cells will gradually develop into the fetus, and the trophectoderm cells will contact the uterine cavity epithelial cells and gradually differentiate into mature trophoblast cells. As trophoblast cells gradually differentiate and develop, they will eventually become one of the more important components of placental tissue unique to mammals. The normal development of embryonic trophoblast during the peri-implantation period is one of the key st...

Claims

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Application Information

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IPC IPC(8): C07K14/47
CPCC07K14/4702
Inventor 薛志刚曾桥吕波纪亚忠
Owner TONGJI UNIV