dsRNA和Aza-CdR转染猪肾细胞的全基因组差异甲基化区域分布特征的比较

doi:10.11843/j.issn.0366-6964.2022.08.031

Abstract

Abstract: This study aimed to screen differential methylation genes and analyze the differential methylation regions specific to the Gene Ontology (GO) by comparing the distribution characteristics of differential methylation peaks in the whole genome of PK15 cells transfected by Poly I:C and Aza-CdR. Firstly, the pig 385 K whole genome promoter and CpG Island methylation chip were used based on MeDIP-chip technology. Three groups of experimental materials (porcine PK15 cells transfected with Poly I:C virus mimic, PK15 cells transfected with methylase inhibitor Aza-CdR, and mock cells without treatment) were analyzed to obtain significantly enriched differential methylation peaks between two experimental groups based on Peak DM Value and Peak Score. Secondly, Gene Ontology (GO) annotation of differential methylation genes was performed to screen for differential methylation regions and differential methylation genes. Finally, the differential methylated region DMR was verified by Bisulfite clone sequencing and mRNA fluorescence quantitative expression test. DNA methylation in the whole genome of porcine kidney cells was mainly distributed in the 5' regulatory region. After comparison between groups, especially P vs. C and A vs. C comparisons, it was found that the distribution characteristics of DNA methylation on the genome were related to the density of CpG island and the location of distance from TSS. DNA methylation in the near promoter region (0-+200 bp) significantly affected gene expression. The effect of Poly I:C on PK15 increased the number of hypomethylation promotors at 200 bp (-200-+500 bp) near TSS, indicating that Poly I:C and Aza-CdR have similar potential demethylation effects. Especially, 669 bp Peak Length CG sites were demethylated in the 10459946-10460615 bp region of the BNIP3L gene on pig chromosome 14. It was revealed that Poly I:C and Aza-CdR did not demethylate in all pig genes, but mainly targeted specific promoters of specific genes, demonstrating that CpG islands of these specific promoters were particularly sensitive to Poly I:C and Aza-CdR.

Key words: Poly I:C, Aza-CdR, porcine kidney cells, MeDIP-chip, differential methylation peaks

CLC Number:

S813.1

WANG Yong, WANG Huaidong, GUO Yongqing, YU Ying, WANG Chuduan, WANG Xiaoshuo. Comparison of Genome-wide Differential Methylation Regions in dsRNA and Aza-CdR Transfected Porcine Kidney Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2739-2750.

References 30

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Comparison of Genome-wide Differential Methylation Regions in dsRNA and Aza-CdR Transfected Porcine Kidney Cells

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