Large Scale Purification of RNA Nanoparticles by Preparative Ultracentrifugation
Purification of large quantities of supramolecular RNA complexes is of paramount importance due to the large quantities of RNA needed and the purity requirements for in vitro and in vivo assays. Purification is generally carried out by liquid chromatography (HPLC), polyacrylamide gel electrophoresis (PAGE), or agarose gel electrophoresis (AGE). Here, we describe an efficient method for the large-scale purification of RNA prepared by in vitro transcription using T7 RNA polymerase by cesium chloride (CsCl) equilibrium density gradient ultracentrifugation and the large-scale purification of RNA nanoparticles by sucrose gradient rate-zonal ultracentrifugation or cushioned sucrose gradient rate-zonal ultracentrifugation.
Key wordsRNA Nanoparticles Ultracentrifugation Nanotechnology Nanobiotechnology RNA nanotechnology RNA therapeutics Large-scale purification
The research was supported by NIH grants R01-EB003730 and U01-CA151648 to P.G. The content is solely the responsibility of the authors and does not necessarily represent the official views of NIH. Funding to Peixuan Guo’s Endowed Chair in Nanobiotechnology position is from the William Fairish Endowment Fund. PG is a cofounder of Kylin Therapeutics, Inc., RNA Nano, LLC., and Biomotor and Nucleic Acid Nanotechnology Development Corp., Ltd.
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