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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
Transcriptome-Wide Analysis of N6-Methyladenosine Modification in the Liver of Geese at Different Growth Stages
Chuan Li1,2, Jintao Wu2, Shuibing Liu2
1College of Life Sciences, Jiangxi Science and Technology Normal University, Nanchang 330013, China.
Abstract:
N6-methyladenosine (m6A) is a reversible RNA modification that dynamically regulates gene expression by modulating RNA stability, splicing, nuclear export, translation, and maturation-thereby orchestrating organismal development. In birds, including geese, the liver is a multi-functional organ central to metabolic regulation. Studies on the dynamic patterns of RNA m6A modifications during healthy liver growth and development remain limited. Here, we performed integrative methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) on liver tissues from geese at three biologically defined stages: post-hatch day 0 (0 week, P), fast growth (10 weeks, F), and sexual maturation (30 weeks, S). The level of m6A modification in total RNA extracted from liver tissues was higher in P than in F samples. Compared with other groups, the S group recorded the lowest m6A modification. In addition, 1641, 668, and 558 m6A peaks were differentially modified in the P, F, and S groups, respectively. The m6A peaks in the liver of the three groups were mainly enriched in the coding sequence and 3' untranslated region. Moreover, integrated multi-omics analysis (MeRIP-seq and RNA-seq), combined with protein-protein interaction networks analysis, identified CDK1 as a core cell cycle regulator and IGF2BP3-a well-established m6A reader-as a consistently differentially expressed gene across all developmental stages. The m6A-regulated cell cycle, p53 signaling pathway, and pyrimidine metabolism pathway were identified in liver tissue as novel potential targets for controlling geese growth and metabolism. Together, these findings shed light on the dynamic regulation of RNA methylation during distinct growth phases in geese and advance our understanding of epigenetic mechanisms underlying poultry liver development.
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