DNA Origami-Enabled Gene Localization of Repetitive Sequences

Abstract

Repetitive sequences, which make up over 50% of human DNA, have diverse applications in disease diagnosis, forensic identification, paternity testing, and population genetic analysis due to their crucial functions for gene regulation. However, representative detection technologies such as sequencing and fluorescence imaging suffer from time-consuming protocols, high cost, and inaccuracy of the position and order of repetitive sequences. Here, we develop a precise and cost-effective strategy that combines the high resolution of atomic force microscopy with the shape customizability of DNA origami for repetitive sequence-specific gene localization. "Tri-block" DNA structures were specifically designed to connect repetitive sequences to DNA origami tags, thereby revealing precise genetic information in terms of position and sequence for high-resolution and high-precision visualization of repetitive sequences. More importantly, we achieved the results of simultaneous detection of different DNA repetitive sequences on the gene template with a resolution of ∼6.5 nm (19 nt). This strategy is characterized by high efficiency, high precision, low operational complexity, and low labor/time costs, providing a powerful complement to sequencing technologies for gene localization of repetitive sequences.

Three-Point-Star Deoxyribonucleic Acid Tiles with the Core Arm Length at Three Half-Turns for Two-Dimensional Archimedean Tilings and Beyond

“…Structural DNA nanotechnology has been intensely investigated for many potential applications such as drug delivery, − biotechnology, − DNA computing, , and nanofabrication. − Generally, DNA nanotechnology involves two continuous architectures: weaving DNA building motifs and tiling the motifs into 0–3D DNA arrays. The essentially woven building motifs include double crossover (DX) and coupled DX (cDX) tiles (both DAE ( d ouble crossover, a ntiparallel, an e ven number of half-turns) and DAO ( d ouble crossover, a ntiparallel, an o dd number of half-turns) tiles are well-known DX tiles), − point-star (abbreviated as PS) tiles, − DNA origami, − and single-stranded tiles and bricks. , The PS tiles each include a looped scaffold strand and several main and auxiliary helper strands, in which the two parts of the tile core and overhangs can be differentiated with the looped scaffold strand and the auxiliary helper strands, respectively (refer to Figure ).…”