HPLC purification of in vitro transcribed long RNA

Abstract

In vitro transcription of DNA with phage RNA polymerases is currently the most ef ficient method to produce long sequence-specific RNA. While the reaction can yield large quantities of RNA, it contains impurities due to various unwanted activities of the polymerases. Here, we described an easily performed HPLC purification that removes multiple contaminants from in vitro transcribed RNA and is scalable. The puri fied RNA is translated at much greater levels, especially in primary cells and in vivo. HPLC purification of RNA containing modi fied nucleosides that suppress RNA-mediated activation of innate immune sensors leads to a non-immunogenic RNA with superior translational capacity. © Springer Science+Business Media New York 2013.