化学修饰的 RNA 纳米颗粒与 siRNA 功能化的共转录组装。
Co-transcriptional assembly of chemically modified RNA nanoparticles functionalized with siRNAs.
机构信息
Computational RNA Structure Group, Center for Cancer Research Nanobiology Program, NCI, Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702, United States.
出版信息
Nano Lett. 2012 Oct 10;12(10):5192-5. doi: 10.1021/nl302302e. Epub 2012 Sep 27.
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.
摘要
我们报道了一种通用的方法，即在 T7 RNA 聚合酶体外转录过程中，一锅法合成化学修饰的功能性 RNA 纳米颗粒。在锰离子存在的情况下，野生型 T7 RNA 聚合酶对 2'-氟-dNTP 的掺入效率显著提高，从而高产化学修饰的 RNA 纳米颗粒，这些颗粒功能化的 siRNA 能够抵抗人血清中的核酸酶。此外，未纯化的转录混合物可用于体外的初步功能实验。
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