Identification of RNAs containing TOP3B cleavage complexes

Human TOP3B is unique as it can act as RNA topoisomerase. For identification of RNAs with TOP3B cleavage sites, we engineered a “self-trapping” mutant of TOP3B (R338W TOP3B) and performed performed native RNA Immunoprecipitation and sequencing without crosslinking (Native RIP seq). 2,856 RNAs (Supplementary Figure 7F) were enriched in R338W TOP3B immunoprecipitated samples and found that TOP3B regulates key cellular RNA-related pathways. Overall design: Native RIP-seq in HEK293 cells transfected with FLAG tagged R338W TOP3B