Song In Hyun, Shin Seung Won, Park Kyung Soo, Lansac Yves, Jang Yun Hee, Um Soong Ho
School of Chemical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do, 440-746, South Korea.
GREMAN, UMR7347, Université François Rabelais, 37200, France.
Sci Rep. 2015 Dec 4;5:17722. doi: 10.1038/srep17722.
With the advent of nanotechnology, a variety of nanoarchitectures with varied physicochemical properties have been designed. Owing to the unique characteristics, DNAs have been used as a functional building block for novel nanoarchitecture. In particular, a self-assembly of long DNA molecules via a piece DNA staple has been utilized to attain such constructs. However, it needs many talented prerequisites (e.g., complicated computer program) with fewer yields of products. In addition, it has many limitations to overcome: for instance, (i) thermal instability under moderate environments and (ii) restraint in size caused by the restricted length of scaffold strands. Alternatively, the enzymatic sewing linkage of short DNA blocks is simply designed into long DNA assemblies but it is more error-prone due to the undeveloped sequence data. Here, we present, for the first time, a comprehensive study for directly combining DNA structures into higher DNA sewing constructs through the 5'-end cohesive ligation of T4 enzyme. Inspired by these achievements, the synthesized DNA nanomaterials were also utilized for effective detection and real-time diagnosis of cancer-specific and cytosolic RNA markers. This generalized protocol for generic DNA sewing is expected to be useful in several DNA nanotechnology as well as any nucleic acid-related fields.
随着纳米技术的出现,人们设计出了各种具有不同物理化学性质的纳米结构。由于DNA具有独特的特性,它已被用作构建新型纳米结构的功能组件。特别是,通过一段DNA短链实现长DNA分子的自组装已被用于构建此类结构。然而,这需要许多苛刻的前提条件(例如,复杂的计算机程序),且产品产量较低。此外,它还有许多需要克服的局限性:例如,(i)在中等环境下热稳定性差,(ii)由于支架链长度受限导致尺寸受限。另外,将短DNA片段进行酶促连接形成长DNA组装体的设计虽简单,但由于序列数据不完善,更容易出错。在此,我们首次展示了一项全面的研究,即通过T4酶的5'-末端粘性连接将DNA结构直接组合成更高阶的DNA连接构建体。受这些成果的启发,合成的DNA纳米材料还被用于癌症特异性和胞质RNA标志物的有效检测和实时诊断。这种通用的DNA连接方案有望在多种DNA纳米技术以及任何与核酸相关的领域中发挥作用。