Co-transcriptional Assembly of Chemically Modified RNA Nanoparticles Functionalized with siRNAs
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- 作者：Kirill A. Afonin ; Maria Kireeva ; Wade W. Grabow ; Mikhail Kashlev ; Luc Jaeger ; Bruce A. Shapiro
- 刊名：Nano Letters
- 出版年：2012
- 出版时间：October 10, 2012
- 年：2012
- 卷：12
- 期：10
- 页码：5192-5195
- 全文大小：286K
- 年卷期：v.12,no.10(October 10, 2012)
- ISSN：1530-6992
文摘
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鈥?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.