Virginia G. Piper Biodesign Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University , Tempe, Arizona 85287, United States.
Department of Bioengineering and Dalton Cardiovascular Research Center, University of Missouri , Columbia, Missouri 65211, United States.
Anal Chem. 2017 Dec 19;89(24):13039-13043. doi: 10.1021/acs.analchem.7b03979. Epub 2017 Dec 4.
Aerolysin protein pore has been widely used for sensing peptides and proteins. However, only a few groups explored this nanopore for nucleic acids detection. The challenge is the extremely low capture efficiency for nucleic acids (>10 bases), which severely lowers the sensitivity of an aerolysin-based genetic biosensor. Here we reported a simple and easy-to-operate approach to noncovalently transform aerolysin into a highly nucleic acids-sensitive nanopore. Through a remote pH-modulation mechanism, we simply lower the pH on one side of the pore, then aerolysin is immediately "activated" and enabled to capture target DNA/RNA efficiently from the opposite side of the pore. This mechanism also decelerates DNA translocation, a desired property for sequencing and gene detection, allowing temporal separation of DNAs in different lengths. This method provides insight into the nanopore engineering for biosensing, making aerolysin applicable in genetic and epigenetic detections of long nucleic acids.
溶细胞素蛋白孔已被广泛用于检测肽和蛋白质。然而，只有少数几个研究小组探索了该纳米孔用于核酸检测。挑战在于核酸（>10 个碱基）的捕获效率极低，这极大地降低了基于溶细胞素的遗传生物传感器的灵敏度。在这里，我们报告了一种简单易用的方法，可将溶细胞素非共价转化为高度核酸敏感的纳米孔。通过远程 pH 调节机制，我们只需在孔的一侧降低 pH 值，然后溶细胞素立即“激活”，并能够从孔的另一侧有效地捕获靶 DNA/RNA。该机制还会减缓 DNA 转位，这是测序和基因检测所期望的特性，可在不同长度的 DNA 之间实现时间分离。该方法为纳米孔工程在生物传感中的应用提供了新的思路，使溶细胞素适用于长核酸的遗传和表观遗传检测。
