A simplified and high-yield purification method for recombinant RNase R

RNase R is a 3'→5' exoribonuclease that selectively degrades linear RNAs. As such, RNase R has become a key reagent for circRNA production and analysis. This is particularly true for the in vitro production of circRNA research tools and therapeutic candidates. Unfortunately, the high cost of commercial RNase R is restrictive. We report an accessible, cost-effective method to purify high-quality recombinant RNase R from E. coli using single-step Ni NTA chromatography on entry level FPLC systems. By achieving complete linear RNA digestion while maintaining circRNA, the resulting enzyme matches performance attributes of commercially available RNase R.