Fang Po-Yu, Bowman Jessica C, Gómez Ramos Lizzette M, Hsiao Chiaolong, Williams Loren Dean
School of Chemistry and Biochemistry, Georgia Institute of Technology Atlanta GA 30332 USA
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology Atlanta GA 30332 USA.
RSC Adv. 2018 Jun 12;8(38):21399-21406. doi: 10.1039/c8ra02084a. eCollection 2018 Jun 8.
Virus Like Particles (VLPs) are devices for RNA packaging, protection and delivery, with utility in fundamental research, drug discovery, and disease treatment. Using for combined expression and packaging of non-viral RNAs into Qβ VLPs, we investigated the extent of chemical protection conferred by packaging of RNA in VLPs. We also probed relationships between packaging efficiency and RNA size, sequence and intrinsic compaction. We observe that VLP packaging protects RNA against assault by small diffusible damaging agents such as hydroxyl radicals and divalent cations. By contrast, the extent of unmediated cleavage, in the absence of reactive species, is the same for RNA that is free or packaged within VLPs, and is very slow. packaging of RNA within VLPs appears to be more efficient for intrinsically compact RNAs, such as rRNA, and less efficient for unstructured, elongated RNA such as mRNA. Packaging efficiency is reduced by addition of the ribosome binding site to a target RNA. The Qβ hairpin is necessary but not sufficient for efficient packaging.
病毒样颗粒(VLPs)是用于RNA包装、保护和递送的工具,在基础研究、药物发现和疾病治疗中具有实用价值。通过使用[具体方法]将非病毒RNA联合表达并包装到Qβ病毒样颗粒中,我们研究了RNA包装在病毒样颗粒中所赋予的化学保护程度。我们还探究了包装效率与RNA大小、序列及内在压缩性之间的关系。我们观察到,病毒样颗粒包装可保护RNA免受诸如羟基自由基和二价阳离子等小分子可扩散损伤剂的攻击。相比之下,在没有反应性物质的情况下,游离或包装在病毒样颗粒内的RNA的非介导切割程度相同,且非常缓慢。对于诸如rRNA等内在紧密的RNA,RNA在病毒样颗粒内的包装似乎更有效,而对于诸如mRNA等无结构的延长RNA则效率较低。将核糖体结合位点添加到靶RNA会降低包装效率。Qβ发夹对于有效包装是必要的,但并不充分。