中文摘要：引入噬菌体MS2病毒样颗粒（VLPs）作为基因和药物传递工具，需要优化其生产和纯化程序。PEG沉淀法可有效纯化VLP，而pH和不同电解质对纯化的MS2 VLP的稳定性、大小和均一性的影响，以及封装的RNA序列仍有待阐明。为此，本研究制备了一种能够产生内含shRNA的VLP的载体，并评估了不同缓冲液/溶液中VLP的大小、多分散指数和保护内含shRNA的能力。研究了含或不含NaNO3的Tris、HEPES和PBS，以及不同pH值和离子浓度下单独使用NaNO3，发现pH为8的100 mM NaNO3 Tris缓冲液是新的最佳MS2 VLP生产缓冲液，能够抑制VLPs聚集。这些VLP的大小为27-30 nm，均一性好，在4℃下至少能维持12个月的稳定性。此外，所得MS2 VLP在类似于体内的条件下具有高效性，并至少稳定48小时。在新引入的缓冲液中生产的MS2 VLP的这些特性使其成为治疗剂递送的合适候选物。
外文摘要：Introducing bacteriophage MS2 virus-like particles (VLPs) as gene and drug delivery tools increases the demand for optimizing their production and purification procedure. PEG precipitation method is used efficiently to purify VLPs, while the effects of pH and different electrolytes on the stability, size, and homogeneity of purified MS2 VLPs, and the encapsulated RNA sequences remained to be elucidated. In this regard, a vector, capable of producing VLP with an shRNA packed inside was prepared. The resulting VLPs in different buffers/solutions were assessed for their size, polydispersity index, and ability to protect the enclosed shRNA. We report that among Tris, HEPES, and PBS, with or without NaNO3, and also NaNO3 alone in different pH and ionic concentrations, the 100 mM NaNO3-Tris buffer with pH:8 can be used as a new and optimal MS2 VLP production buffer, capable of inhibiting the VLPs aggregation. These VLPs show a size range of 27-30 nm and suitable homogeneity with minimum 12-month stability at 4 degrees C. Moreover, the resulting MS2 VLPs were highly efficient and stable for at least 48 h in conditions similar to in vivo. These features of MS2 VLPs produced in the newly introduced buffer make them an appropriate candidate for therapeutic agents' delivery.
 
外文关键词：MICRORNA DELIVERY-SYSTEM；CELL-SPECIFIC DELIVERY；MESSENGER-RNA；BACTERIOPHAGE-MS2；SURFACE；AGGREGATION；STABILITY；PROTEINS；CAPSIDS；IMPACT
作者：Hashemi, K；Seno, MMG；Ahmadian, MR；Malaekeh-Nikouei, B；Bassami, MR；Dehghani, H；Afkhami-Goli, A
作者单位：Ferdowsi Univ Mashhad；Heinrich Heine Univ；Hosp Sick Children；Mashhad Univ Med Sci
期刊名称：SCIENTIFIC REPORTS
期刊影响因子：3.998
出版年份：2021
出版刊次：1
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