Sequence‐Guided Localization of DNA Hybridization Enables Highly Selective and Robust Genotyping

Genetic variations are always related to human diseases or susceptibility to therapies. Nucleic acid probes that precisely distinguish closely related sequences become an indispensable requisite both in research and clinical applications. Here, a Sequence‐guided DNA LOCalization for leaKless DNA detection (SeqLOCK) is introduced as a technique for DNA hybridization, where the intended targets carrying distinct “guiding sequences” act selectively on the probes. In silicon modeling, experimental results reveal considerable agreement (R2 = 0.9228) that SeqLOCK is capable of preserving high discrimination capacity at an extraordinarily wide range of target concentrations. Furthermore, SeqLOCK reveals high robustness to various solution conditions and can be directly adapted to nucleic acid amplification techniques (e.g., polymerase chain reaction) without the need for laborious pre‐treatments. Benefiting from the low hybridization leakage of SeqLOCK, three distinct variations with a clinically relevant mutation frequency under the background of genomic DNA can be discriminated simultaneously. This work establishes a reliable nucleic acid hybridization strategy that offers great potential for constructing robust and programmable systems for molecular sensing and computing. In this work, the mechanism of sequence‐guided DNA localization is introduced. Due to the close proximity by the hybridization of distinct guiding sequence, the reaction between the target and the corresponding probe can be selectively accelerated. Experimental results show that the spurious reactions by excessive concentrations of targets can be kinetically constrained, resulting in high selectivity and robustness.