Abstract: The present disclosure provides methods of reducing, minimizing, or inhibiting the formation of double-stranded ribonucleic acid (dsRNA) during the preparation of ribonucleic acid (RNA), such as messenger ribonucleic acid (mRNA), by adding at least one chaotropic agent to a starting reaction mixture. The present disclosure provides methods of reducing, minimizing, or inhibiting intramolecular base-pairing within a ribonucleic acid (RNA) transcript and/or intermolecular base-pairing between an RNA transcript and deoxyribonucleic acid (DNA) or another RNA during the preparation of ribonucleic acid (RNA), by adding at least one chaotropic agent to a starting reaction mixture.

Abstract: Disclosed herein are high Tm RNA nanostructures that can be composed of one or more modules or motifs to build RNA nanostructures with or without layers. The RNA nanostructures can have a core domain and three or more double-stranded arms and formulations thereof to conjugate high copy numbers of therapeutics, pH responsive or enzyme cleavable drug cargo. Also described herein is a design strategy for generation of synthetic RNA oligonucleotides that can self assemble into highly thermostable RNA structures. Also described herein are uses of the RNA nanostructures described herein.