Jasinski Daniel L, Khisamutdinov Emil F, Lyubchenko Yuri L, Guo Peixuan
ACS Nano. 2014 Aug 26;8(8):7620-9. doi: 10.1021/nn502160s.
Recent advances in RNA nanotechnology allow the rational design of various nanoarchitectures. Previous methods utilized conserved angles from natural RNA motifs to form geometries with specific sizes. However, the feasibility of producing RNA architecture with variable sizes using native motifs featuring fixed sizes and angles is limited. It would be advantageous to display RNA nanoparticles of diverse shape and size derived from a given primary sequence. Here, we report an approach to construct RNA nanoparticles with tunable size and stability. Multifunctional RNA squares with a 90° angle were constructed by tuning the 60° angle of the three-way junction (3WJ) motif from the packaging RNA (pRNA) of the bacteriophage phi29 DNA packaging motor. The physicochemical properties and size of the RNA square were also easily tuned by modulating the “core” strand and adjusting the length of the sides of the square via predictable design. Squares of 5, 10, and 20 nm were constructed, each showing diverse thermodynamic and chemical stabilities. Four “arms” extending from the corners of the square were used to incorporate siRNA, ribozyme, and fluorogenic RNA motifs. Unique intramolecular contact using the pre-existing intricacy of the 3WJ avoids relatively weaker intermolecular interactions via kissing loops or sticky ends. Utilizing the 3WJ motif, we have employed a modular design technique to construct variable-size RNA squares with controllable properties and functionalities for diverse and versatile applications with engineering, pharmaceutical, and medical potential. This technique for simple design to finely tune physicochemical properties adds a new angle to RNA nanotechnology.
RNA纳米技术的最新进展使得各种纳米结构的合理设计成为可能。以往的方法利用天然RNA基序的保守角度来形成具有特定尺寸的几何形状。然而，利用具有固定尺寸和角度的天然基序生产具有可变尺寸的RNA结构的可行性是有限的。展示源自给定一级序列的不同形状和尺寸的RNA纳米颗粒将是有利的。在这里，我们报告了一种构建具有可调尺寸和稳定性的RNA纳米颗粒的方法。通过调整噬菌体phi29 DNA包装马达的包装RNA（pRNA）的三向接头（3WJ）基序的60°角，构建了具有90°角的多功能RNA正方形。RNA正方形的物理化学性质和尺寸也可以通过调节“核心”链并通过可预测的设计调整正方形边的长度来轻松调节。构建了5、10和20纳米的正方形，每个都显示出不同的热力学和化学稳定性。从正方形的角延伸出的四个“臂”用于掺入siRNA、核酶和荧光RNA基序。利用3WJ预先存在的复杂性进行独特的分子内接触，避免了通过亲吻环或粘性末端的相对较弱的分子间相互作用。利用3WJ基序，我们采用模块化设计技术构建了具有可控性质和功能的可变尺寸RNA正方形，用于具有工程、制药和医学潜力的各种通用应用。这种简单设计以微调物理化学性质的技术为RNA纳米技术增添了新的视角。
