Transcriptome analysis of the effect of enrichment and regulation of m6A by synthetic writer and reader in human cells

To investigate the basic principles of epigenetic control, we established an orthogonal epigenetic regulatory system in mammalian cells using N6-methyladenine (m6A), a DNA modification not commonly found in metazoan epigenomes. We developed a synthetic initiator module (synI) capable of establishing m6A marks in a sequence-specific manner at designer reporter loci integrated in the human genome, and a synthetic readout module (synR) which selectively reads m6A and consequently induces transcriptional changes. To ensure our orthogonal m6A system has minimal interaction with endogenous human transcriptional machineries, we performed RNA-sequencing analysis and determined that both synI and synR expression have minimal effect on 293FT transcriptome.