Analysis of m6A methylome in CHAPIR overexpressed heart [MeRIP]

To elucidate the molecular mechanism by which cardiac-hypertrophy-associated piRNA (CHAPIR) regulates m6A modification, we performed m6A methylated RNA immunoprecipitation sequencing (MeRIP-seq) in control or CHAPIR-overexpressing mice heart. The sequential analysis of m6A peaks showed that RGACH motif was highly enriched within m6A sites in heart and that is aligning with the classical consensus sequence of mammals ‘RGACH’, where ‘R’ indicates purine (A/G) and ‘H’ indicates non-guanine base (A/C/U). In CHAPIR treated heart, m6A mostly occurred in mRNAs (89.5%) and only about 10.5% were identified in non-coding RNAs. The majority of mRNAs contain one or two m6A peaks (89.2%) and 10.8% mRNA contain >2 m6A peaks . M6A peaks were predominantly distributed in coding sequences (CDSs), 3’ untranslated regions (3’UTRs) and near stop codon. Total RNAs were extracted from CHAPIR mimic or mimic-negative control (NC) overexpressed heart. MeRIP was performed to enrich m6A methylated mRNAs using anti-N6-methyadenosine (m6A) antibody. We used commercial kit (KAPA Stranded mRNA-seq Kit) for RNA-seq library preparation of both m6A enriched RNAs and input mRNAs. Then, high throughput RNA sequencing was performed to identify differences in the gene expression profile. Please note that each processed data was generated from both IP and input samples, and is linked to the corresponding IP sample records.