School of Chemistry and Chemical Engineering, School of Mechanical and Electrical Engineering, Guangzhou University, Guangzhou, Guangdong, 510006, China.
School of Life Sciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou, China.
Analyst. 2023 Sep 11;148(18):4346-4355. doi: 10.1039/d3an01126g.
Glass nanopore is an ideal candidate for biosensors due to its unique advantages such as label-free analysis, single-molecule sensitivity, and easy operation. Previous studies have shown that glass nanopores can distinguish different lengths of double-stranded DNA (dsDNA) at the same time with the length-resolution ability. Based on this, we proposed a novel design of a dsDNA block containing a programmable sensing site inside, which can be programmed to respond to different target molecules and cleaved into two smaller DNA blocks. When programming the sensing site with different sequences, for example, programming it as the substrate of GR-5 DNAzyme and CRISPR-Cas12a system, the DNA block could realize Pb and cfDNA detection with the length-resolution ability of the glass nanopore. This strategy achieved a Pb detection range from 0.5 nM to 100 nM, with a detection limit of 0.4 nM, and a BRCA-1 detection range from 1 pM to 10 pM, with a detection limit of 1 pM. The programable sensing site is easy to design and has strong expandability, which gives full play to the advantages of glass nanopore in length-resolution ability for dsDNA, and is expected to become an optional design for biosensing strategy for the glass nanopore as a biosensing platform.
玻璃纳米孔由于其无标记分析、单分子灵敏度和易于操作等独特优势，是生物传感器的理想候选者。先前的研究表明，玻璃纳米孔具有长度分辨率能力，可同时区分不同长度的双链 DNA（dsDNA）。基于此，我们提出了一种新型 dsDNA 块的设计，其中包含可编程的传感位点，可被编程以响应不同的靶分子并切割成两个较小的 DNA 块。当用不同的序列对传感位点进行编程时，例如将其编程为 GR-5 DNA 酶和 CRISPR-Cas12a 系统的底物时，DNA 块可以利用玻璃纳米孔的长度分辨率能力实现 Pb 和 cfDNA 的检测。该策略实现了从 0.5 nM 到 100 nM 的 Pb 检测范围，检测限为 0.4 nM，以及从 1 pM 到 10 pM 的 BRCA-1 检测范围，检测限为 1 pM。可编程传感位点易于设计且具有很强的可扩展性，充分发挥了玻璃纳米孔在 dsDNA 长度分辨率能力方面的优势，有望成为玻璃纳米孔作为生物传感平台的生物传感策略的可选设计。
