RNA sequencing of H9c2 cells upon transfection of inosine-containing RNA and treatment with inosine nucleoside

Inosine monophosphate (IMP) is a precursor in the synthesis of AMP and GMP, but has the potential to become phosphorylated forming ITP or can be formed via the deamination of ATP. Once in triphosphate form, ITP can be misincorporated into RNA during transcription potentially affecting the function of the nascent RNA. The ubiquitous enzyme ITPase is found in all 3 domains of life and functions to dephosphorylate ITP, among other (d)NTPs, from the nucleotide pool preventing misincorporation. Loss of ITPase leads to a fatal multisystem disorder with epileptic encephalopathy in humans. Illumina RNA sequencing and direct RNA nanopore sequencing were used to both evaluate the cellular response upon transfection of inosine-containing RNA and detect inosine misincorporation following treatment with inosine nucleoside in H9c2 cells.