The effects of RNA binding activity and acetyltransferase enzyme activity of NAT10 on heart development

The goal of this study is to investigate whether RNA binding activity and acetyltransferase enzyme activity of NAT10 regulates heart development Overall design: At day 1 after birth, Nat10flox/flox mice were injected with AAV9-cTnT-GFP (1X 1011 VP/mouse) subcutaneously, and Nat10-CKO mice were injected with equal amount of AAV9-cTnT-GFP, AAV9-cTnT-NAT10, AAV9-cTnT-NAT10G641E, and AAV9-cTnT-NAT10K209A, respectively. Mice were sacrificed at day 12-14 after injection. Heart tissues were collected. mRNA profiles in these heart tissues were generated by deep sequencing using Illumina Novaseq 6000 platform (n=3 for each group). Methods: Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany) from heart tissues. Three independent biological replicates for each group were used for RNA-seq. RNA-seq was performed by deep sequencing using an Illumina Novaseq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome(GRCm38/mm10) with Hisat2 v2.0.5, and the aligned reads were used to quantify mRNA expression by using featureCounts v1.5.0-p3. Our study represents the first detailed analysis of the effects on heart transcriptomes by the RNA binding activity and acetyltransferase enzyme activity of NAT10, generated by RNA-seq technology.