Haque Farzin, Xu Congcong, Jasinski Daniel L, Li Hui, Guo Peixuan
Division of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, The Ohio State University, Columbus, OH, 43210, USA.
Department of Physiology and Cell Biology, College of Medicine, The Ohio State University, Columbus, OH, 43210, USA.
Methods Mol Biol. 2017;1632:359-380. doi: 10.1007/978-1-4939-7138-1_23.
RNA is rapidly emerging as a versatile building block for nanoparticle assembly due to its simplicity in base pairing, while exhibiting diversity in function such as enzymatic activity similar to some proteins. Recent advances in RNA nanotechnology have generated significant interests in applying RNA nanoparticles for various applications in nanotechnology and nanomedicine. In particular, assessing the effect of size and shape on cell entry and intracellular trafficking as well as in vivo biodistribution of nanoparticles is challenging due to the lack of nanoparticles rich in structure while varying in size and shape. RNA nanotechnology exemplified by the packaging RNA (pRNA) of bacteriophage phi29 DNA packaging motor has provided a different prospect in nanoparticle designs. Of note, there is a robust three-way junction (3WJ) motif in pRNA which can serve as an adaptable scaffold to construct thermodynamically stable 2D planar and 3D globular RNA architectures with tunable shapes and sizes, and harboring various targeting, therapeutic, and imaging modules. This chapter focuses on the methods for constructing pRNA-3WJ based nanoparticles with controllable sizes and shapes, and assessment of their biodistribution profiles in cancer mouse models after systemic injection and ocular mouse models following subconjunctival injection.
由于RNA在碱基配对方面的简单性，它正迅速成为纳米颗粒组装的一种多功能构建模块，同时在功能上表现出多样性，如具有与某些蛋白质相似的酶活性。RNA纳米技术的最新进展引发了人们对将RNA纳米颗粒应用于纳米技术和纳米医学各种应用的浓厚兴趣。特别是，由于缺乏结构丰富但尺寸和形状各异的纳米颗粒，评估尺寸和形状对细胞摄取、细胞内运输以及纳米颗粒在体内生物分布的影响具有挑战性。以噬菌体phi29 DNA包装马达的包装RNA（pRNA）为例的RNA纳米技术在纳米颗粒设计方面提供了不同的前景。值得注意的是，pRNA中存在一个强大的三向连接（3WJ）基序，它可以作为一个适应性支架，构建具有可调形状和尺寸的热力学稳定的二维平面和三维球状RNA结构，并承载各种靶向、治疗和成像模块。本章重点介绍构建具有可控尺寸和形状的基于pRNA-3WJ的纳米颗粒的方法，以及在全身注射后在癌症小鼠模型和结膜下注射后在眼部小鼠模型中评估其生物分布情况。
