title: Co-transcriptional assembly of chemically modified RNA nanoparticles functionalized with siRNAs.
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.
authors.affiliation: Computational RNA Structure Group, Center for Cancer Research Nanobiology Program, NCI, Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702, United States.
authors.forename: Kirill A
authors.initials: KA
authors.lastname: Afonin
authors.forename: Maria
authors.initials: M
authors.lastname: Kireeva
authors.forename: Wade W
authors.initials: WW
authors.lastname: Grabow
authors.forename: Mikhail
authors.initials: M
authors.lastname: Kashlev
authors.forename: Luc
authors.initials: L
authors.lastname: Jaeger
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Shapiro
mesh_terms: D045744:Cell Line, Tumor; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D006801:Humans; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D036103:Nanotechnology; D012313:RNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D014764:Viral Proteins; D012313:RNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases