Entropy-Driven Three-Dimensional DNA Nanofireworks for Simultaneous Real-Time Imaging of Telomerase and MicroRNA in Living Cells

Abstract

Real-time monitoring of different types of intracellular tumor-related biomarkers is of key importance for the identification of tumor cells. However, it is hampered by the low abundance of biomarkers, inefficient free diffusion of reactants, and complex cytoplasmic milieu. Herein, we present a stable and general method for in situ imaging of microRNA-21 and telomerase utilizing simple highly integrated dual tetrahedral DNA nanostructures (TDNs) that can naturally enter cells, which could initiate to form the three-dimensional (3D) higher-order DNA superstructures (DNA nanofireworks, DNFs) through a reliable target-triggered entropy-driven strand displacement reaction in living cells for remarkable signal amplification. Importantly, the excellent biostability, biocompatibility, and sensitivity of this approach benefited from (i) the precise multidirectional arrangement of probes with a pure DNA structure and (ii) the local target concentration enhanced by the spatially confined microdomain inside the DNFs. This strategy provides a pivotal molecular toolbox for broad applications such as biomedical imaging and early precise cancer diagnosis.

Accelerated Hybridization Chain Reaction Kinetics Using Poly DNA Tetrahedrons and Its Application in Detection of Aflatoxin B1

“…To prepare the pTDNs-HCR, we first synthesized TDN-A and TDN-B through mixing corresponding sequences (Table S1) in TM buffer (20 mM Tris-HCl, 50 mM MgCl 2 , 50 mM KCl, pH = 8.0), 36,37 and the mixture was heated to 95 °C for 5 min and rapidly cooled to 4 °C. 38 Afterward, equal concentrations of TDN-A and TDN-B were mixed for 1 h at 37 °C to generate pTDNs. Finally, the solutions above were blended with H1 and H2 under room temperature for 0.5 h to form a pTDNs-HCR structure.…”

Integration of Isothermal Enzyme‐Free Nucleic Acid Circuits for High‐Performance Biosensing Applications

“…For example, He et al proposed a miRNA-triggered entropy-driven three-dimensional DNA amplification (EDTD) strategy in living cells. [21][22] Through the interaction of target miRNAs with EDTD, a spontaneous DNA reaction was specifically initiated in an entropy-driven manner. The design improves biological stability and endocytosis efficiency by a unique DNA tetrahedral framework structure compared to free DNA.…”

Efficient Three-Dimensional DNA Nanomachine Guided by a Robust Tetrahedral DNA Nanoarray Structure for the Rapid and Ultrasensitive Electrochemical Detection of Matrix Metalloproteinase 2

“…The majority of the traditional 3D DNA nanomachines not only moved on the disordered DNA tracks composed of two-dimensional (2D) or one-dimensional (1D) single-stranded (ss)­DNA probes, , but also walked in a ″step-by-step″ fashion with a limited number and length of arms, , which readily led to a deviation from the designed tracks so as to greatly limit the amplification efficiency and slow down the walking rate of the 3D DNA nanomachines. Surprisingly, it was well known that the tetrahedral DNA (TDN) structure has the merits of a predesigned size, structural stability, and excellent biocompatibility. − Hence, it was prospective to make the 3D DNA nanomachine walk with better directionality and a faster reaction rate by using TDNs as rolling and amplification scaffolds.…”