Co-transcriptional Assembly
of Chemically Modified
RNA Nanoparticles Functionalized with siRNAs
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Abstract
We report a generalized methodology for the one-pot production
of chemically modified functional RNA nanoparticles during in vitro
transcription with T7 RNA polymerase. The efficiency of incorporation
of 2′-fluoro-dNTP in the transcripts by the wild type T7 RNA
polymerase dramatically increases in the presence of manganese ions,
resulting in a high-yield production of chemically modified RNA nanoparticles
functionalized with siRNAs that are resistant to nucleases from human
blood serum. Moreover, the unpurified transcription mixture can be
used for functional ex vivo pilot experiments