Danilevich V N, Sorokin V V, Moiseev Ya P, Sizova S V
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia.
Vinogradsky Institute of Microbiology, Federal Research Center Fundamentals of Biotechnology, Russian Academy of Sciences, Moscow, 117312, Russia.
Dokl Biochem Biophys. 2018 Mar;479(1):118-122. doi: 10.1134/S1607672918020187. Epub 2018 May 19.
The patterns of formation of RNA nanoparticles (NPs) during thermal cycling of bacterial total tRNA in the presence of cations Ca, Mn, Ni, Zn, Co, and Cu were studied. The optimal conditions for the production of NPs were found, and it was revealed that their size depends on the ratio of the concentrations of Me and tRNA. The concentration of reagents for obtaining NPs of small size (from 5 to 100 nm) was selected. It was shown that tRNA-based nanoparticles can comprise short (20-50 nt) ribooligonucleotides, including aptamers and siRNAs. The stability of NPs during storage in buffer solutions of various composition was studied. It was found that the initial suspensions of NPs are quite stable, but they are rapidly destroyed in PBS buffer (pH 7.4). A simple and effective stabilizer (polyarginine) was found, the additives of which ensure the preservation of nanoparticles in PBS buffer for more than 5 h. Nanoparticles modified with the stabilizer are resistant to blood serum nucleases and can be used for transfection.
研究了在阳离子钙、锰、镍、锌、钴和铜存在下，细菌总tRNA热循环过程中RNA纳米颗粒（NPs）的形成模式。发现了产生NPs的最佳条件，并且揭示了它们的大小取决于金属离子（Me）和tRNA浓度的比例。选择了用于获得小尺寸（5至100nm）NPs的试剂浓度。结果表明，基于tRNA的纳米颗粒可以包含短（20 - 50个核苷酸）的核糖寡核苷酸，包括适体和小干扰RNA（siRNAs）。研究了NPs在各种组成的缓冲溶液中储存期间的稳定性。发现NPs的初始悬浮液相当稳定，但它们在PBS缓冲液（pH 7.4）中会迅速被破坏。发现了一种简单有效的稳定剂（聚精氨酸），其添加剂可确保纳米颗粒在PBS缓冲液中保存超过5小时。用该稳定剂修饰的纳米颗粒对血清核酸酶具有抗性，可用于转染。
