Identification of mRNAs associated with NAT10 in mouse hearts

The goal of this study is to identify mRNAs associated with NAT10 in mouse hearts. At day 1 after birth, WT mice were injected with AAV9-cTnT-FLAG-NAT10 (3X 1011 VP/mouse) subcutaneously. Mice were sacrificed at day 20 after injection. RNA immunoprecipitation sequence (RIP-seq) assay was performed to identify mRNAs associated with NAT10 in mouse hearts. Then, immunoprecipitated RNA or Input was used for library construction with NEB NextR Ultra™ RNA Library Prep Kit (New England Biolabs). Library quality was assessed on the Agilent Bioanalyzer 2100 system. The libraries were sequenced on an Illumina Novaseq 6000 platform with a paired-end read length of 150 bp according to the standard protocols. Raw data (raw reads) of fastq format were firstly processed using fastp software. In this step, clean data (clean reads) were obtained by removing reads containing adapter, reads containing ploy-N and low- quality reads from raw data. At the same time, Q20, Q30 and GC content of the clean data were calculated. All the downstream analyses were based on the clean data with high quality. Index of the reference genome ( grcm39-110) was built using BWA (v 0.7.12) and clean reads were aligned to the reference genome using BWA mem (v 0.7.12). mRNAs associated with NAT10 in mouse hearts were characterized.