Method for studying mechanism of combined effect of Wnt signal and histone on target gene during differentiation of PGCs

A histone and target gene technology, applied in biochemical equipment and methods, genetic engineering, plant gene improvement, etc., can solve the problem of lack of Wnt signaling pathway and epigenetic factors to coordinate the regulation of target gene transcription.

Pending Publication Date: 2019-10-18
YANGZHOU UNIV
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Problems solved by technology

However, there is currently no method to study whether the Wnt signaling pathway and epigenetic factors cooperate to regulate the transcription of target genes.

Method used

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  • Method for studying mechanism of combined effect of Wnt signal and histone on target gene during differentiation of PGCs
  • Method for studying mechanism of combined effect of Wnt signal and histone on target gene during differentiation of PGCs
  • Method for studying mechanism of combined effect of Wnt signal and histone on target gene during differentiation of PGCs

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Embodiment 1

[0163] The inventor's research shows that the target gene of Wnt / β-catenin / Tcf7l2 is Lin28A / B, based on this, the participation of histone methylation in this regulation process is studied.

[0164] 1. Research on the regulation of Lin28A / B transcription by H3K4me2

[0165] (1) In vitro verification that H3K4me2 positively regulates Lin28A / B transcription

[0166] The inventors have shown that the histone methylase MLL2 (also known as Kmt2d) and the histone demethylase LSD1 can regulate the methylation status of H3K4me2, respectively using the constructed MLL2 and LSD1 lentiviral interference vectors (shMLL2 and shLSD1 lentiviral vector) for follow-up experiments, and the construction method refers to the paper "Research on the Mechanism of Histone H3K4me2 on the Formation of Chicken SSCs" (He Nana. Research on the Mechanism of Histone H3K4me2 on the Formation of Chicken SSCs [D]. 2018. ) method, and the lentiviral packaging was completed by Shanghai Jiman Company.

[0167] ...

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Abstract

The invention provides a method for studying a mechanism of combined effect of a Wnt signal and histone on a target gene during differentiation of PGCs, which relates to the technical field of biology. The method confirms whether histone has a target gene regulating a Wnt signaling pathway by in-vitro and in-vivo experiments, and analyzes whether the histones in ESCs, PGCs and SSCs are differentially enriched in a promoter region of the target gene, and detects whether histone methylation / demethylation affects the response of the target gene to the Wnt signal, and detects the effect of histonedemethylation modifying enzyme on the binding ability of beta-Catenin to a target gene promoter binding site; and according to the detection results, the mechanism of action of the Wnt signal with histones on the target gene during PGCs differentiation is determined. The method is used to study the combined effect of the Wnt signal and histone on the target gene during the formation of PGCs, which is beneficial to understand a network mechanism for the formation of PGCs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for studying the mechanism of Wnt signal and histone acting on target genes in the course of PGCs differentiation. Background technique [0002] It is well known that the regulation of target gene transcription by β-Catenin and TCFs in the Wnt signaling pathway is mainly achieved by changing the state of chromatin, and chromatin is a nucleosome composed of histones in the nucleus and tightly wound DNA on it. formed by compression. The dynamic modification of histones and DNA (epigenetic modification) affects the structural density of chromatin and also affects the transcriptional state of genes, so we have reason to believe that WNT signaling may cooperate with epigenetic factors to regulate the transcription of target genes. However, there is currently no method to study whether the Wnt signaling pathway and epigenetic factors cooperate to regulate the transcription of tar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869G01N33/68C12N15/867
CPCC12Q1/6869G01N33/68C12N15/86C12N2740/15043
Inventor 李碧春张晨左其生王曼金晶李婷婷张亚妮孙红艳
Owner YANGZHOU UNIV
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