Method for researching target gene regulated and controlled by histone methylation in chicken SSCs

A technology of histone methylation and target genes, applied in biochemical equipment and methods, genetic engineering, plant gene improvement, etc., to achieve the effect of simple and reliable research, simple and feasible methods, and clear procedures

Pending Publication Date: 2020-10-30
YANGZHOU UNIV
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Problems solved by technology

However, there is no report on the transcriptional regulation mechanism of histone methylation in chicken spermatogonial stem cells

Method used

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Experimental program
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Embodiment Construction

[0014] Infection of chicken spermatogonial stem cells with lentivirus interference vector

[0015] Chicken spermatogonial stem cells (MOI=15) were infected with lentivirus shRNA-Mll2 and shRNA-Lsd1, and 6 µg / mL polybrene was added to the virus solution. After 48 hours of infection, the medium containing lentivirus was aspirated and replaced with fresh spermatogonial stem cell medium. After continuing to culture for 24-48h, collect RNA samples. After the RNA samples from three independent experiments were mixed, they were sent to Beijing Novogene Technology Co., Ltd. for transcript analysis, and Agilent 2100 bioanalyzer was used for quality detection and sequencing of the purified total cellular RNA.

[0016] Principal component analysis and heat map analysis of sequencing results after interference with Mll2 and Lsd1

[0017] Principal component analysis was performed on the data obtained from the sequencing analysis, and it was found that after interference with Mll2 / Lsd1, ...

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Abstract

The invention discloses a method for regulating a target gene by histone methylation in chicken SSCs, and belongs to the technical field of biology. According to the method, bioinformatics and experimental verification are combined, RNA-seq and CHIP-seq are combined, and key target genes regulated and controlled by histone methylation in chicken SSCs are selected out through screening. The research on the histone methylation regulated target gene is simple and reliable, the conservative property of the target gene is considered, and a theoretical basis is provided for broadening the research of histone methylation on SSCs.

Description

technical field [0001] The invention relates to a method for target genes regulated by histone methylation in chicken SSCs, and belongs to the field of biotechnology. Background technique [0002] In mice, H3K4me3, H3K9me3, and H3K27me3 have been found to exist in SSCs at the same time, but the three modifications have specific regulation methods. H3K4me3 activates gene expression, while H3K9me3 and H3K27me3 inhibit gene expression; overexpression of H3K4 demethylase Lsd1, It will destroy the histone methylation level during spermatogenesis and affect the health of offspring. However, at present, the research on the transcriptional regulation mechanism of histone methylation in chicken spermatogonial stem cells has not been reported, and many unknown target genes regulated by histone methylation have yet to be discovered. [0003] Sperm quality and quantity are crucial for fertilization and embryonic development in humans and animals. The spermatogonial stem cells are the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867C12Q1/6888C12Q1/6869
CPCC12N5/061C12N15/86C12Q1/6888C12Q1/6869C12Q2600/158C12N2510/00C12N2740/15043
Inventor 李碧春张晨左其生张亚妮孙红艳
Owner YANGZHOU UNIV
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