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Methods Mol Biol. 2017;1632:91-105. doi: 10.1007/978-1-4939-7138-1_6.

Cotranscriptional Production of Chemically Modified RNA Nanoparticles.

Author information

1
RNA Biology Laboratory, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA. kireevam@mail.nih.gov.
2
RNA Biology Laboratory, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA.
3
Nanoscale Science Program, Department of Chemistry, University of North Carolina at Charlotte, 9201 University City Boulevard, Charlotte, NC, USA.
4
The Center for Biomedical Engineering and Science, University of North Carolina at Charlotte, Charlotte, NC, USA.
5
RNA Structure and Design Section, RNA Biology Laboratory, National Cancer Institute, National Institutes of Health, Frederick, MD, 21702, USA.

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

PMID:
28730434
DOI:
10.1007/978-1-4939-7138-1_6
[Indexed for MEDLINE]

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