Detecting cytosine-5 methylation in RNA

We generated a map of NSUN6-dependent RNA 5 methyl-cytosine in the human transcriptome by applying RNA bisulfite conversion and sequencing to two sets of cell lines (HUES9 and HEK293) where the RNA cytosine 5 methyl-transfearse NSUN6 has been either knocked out by CRISPR Cas9 or overexpressed using a PiggyBac vector. We apply this and other molecular methods to validate NSUN6 RNA methylation targets with single nucelotide resolution in the human transcriptome. Overall design: Whole transcriptome bisulfite sequencing of wildtype, Nsun6 -/-, Nsun6 over-expresson and Nsun6 rescue human H9 and/or HEK cells; four replicates per sample.