Stewart Jaimie Marie, Subramanian Hari K K, Franco Elisa
Department of Bioengineering, University of California, 900 University Avenue, Riverside, CA 92521, USA.
Department of Mechanical Engineering, University of California, 900 University Avenue, Riverside, CA 92521, USA.
Nucleic Acids Res. 2017 May 19;45(9):5449-5457. doi: 10.1093/nar/gkx063.
Rational design of nucleic acid molecules yields self-assembling scaffolds with increasing complexity, size and functionality. It is an open question whether design methods tailored to build DNA nanostructures can be adapted to build RNA nanostructures with comparable features. Here we demonstrate the formation of RNA lattices and tubular assemblies from double crossover (DX) tiles, a canonical motif in DNA nanotechnology. Tubular structures can exceed 1 μm in length, suggesting that this DX motif can produce very robust lattices. Some of these tubes spontaneously form with left-handed chirality. We obtain assemblies by using two methods: a protocol where gel-extracted RNA strands are slowly annealed, and a one-pot transcription and anneal procedure. We identify the tile nick position as a structural requirement for lattice formation. Our results demonstrate that stable RNA structures can be obtained with design tools imported from DNA nanotechnology. These large assemblies could be potentially integrated with a variety of functional RNA motifs for drug or nanoparticle delivery, or for colocalization of cellular components.
核酸分子的合理设计产生了具有越来越高复杂性、尺寸和功能性的自组装支架。专门用于构建DNA纳米结构的设计方法是否能够适用于构建具有类似特征的RNA纳米结构，这仍是一个悬而未决的问题。在此，我们展示了由双交叉（DX）基元形成的RNA晶格和管状组装体，DX基元是DNA纳米技术中的一个典型基序。管状结构的长度可以超过1μm，这表明这种DX基序能够产生非常坚固的晶格。其中一些管子会自发地形成左手螺旋性。我们通过两种方法获得组装体：一种是将凝胶提取的RNA链缓慢退火的方案，另一种是一锅法转录和退火程序。我们确定了片段切口位置是晶格形成所需的结构条件。我们的结果表明，利用从DNA纳米技术引入的设计工具可以获得稳定的RNA结构。这些大型组装体可能潜在地与各种功能性RNA基序整合，用于药物或纳米颗粒递送或细胞成分的共定位。
