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Using Lipid Nanoparticles for the Delivery of Chemically Modified mRNA into Mammalian Cells

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Cotranscriptional Production of Chemically Modified RNA Nanoparticles

Maria L Kireeva¹, Kirill A Afonin^2,3,4, Bruce A Shapiro^2,5

¹RNA Biology Laboratory, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA. kireevam@mail.nih.gov.

Methods in Molecular Biology (Clifton, N.J.)

|July 22, 2017

Abstract:

RNA nanoparticles consisting of multiple RNA strands of different sequences forming various three-dimensional structures emerge as promising carriers of siRNAs, RNA aptamers, and ribozymes. In vitro transcription of a mixture of dsDNA templates encoding all the subunits of the RNA nanoparticle may result in cotranscriptional self-assembly of the nanoparticle. Based on our experience with production of RNA nanorings, RNA nanocubes, and RNA three-way junctions, we propose a strategy for optimization of the cotranscriptional production of chemically modified ribonuclease-resistant RNA nanoparticles.

Keywords:

2′-F-dCTP 2′-F-dUTP Cotranscriptional assembly In vitro transcription RNA nanocubes RNA nanorings T7 R&DNA polymerase T7 RNA polymerase

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