Structural insights into the AFB1 aptamer coupled with a rationally designed CRISPR/Cas12a-Exo III aptasensor for AFB1 detection

适体 范德瓦尔斯力 化学 分子识别 清脆的 纳米技术 合理设计 分子 材料科学 生物 分子生物学 生物化学 基因 有机化学
作者
Pengfei Ma,Hualin Guo,Hua Ye,Yin Zhang,Zhouping Wang
出处
期刊:International Journal of Biological Macromolecules [Elsevier BV]
卷期号:225: 1164-1171 被引量:30
标识
DOI:10.1016/j.ijbiomac.2022.11.177
摘要

Aflatoxin B1 (AFB1) is a typical food contaminant. A truncated DNA aptamer of AFB1 was reported by our team in previous work. However, the recognition mechanism between aptamer and AFB1 was lacking, which was crucial for the design of related aptasensor. Herein, the binding of aptamer to AFB1 was systematically studied and found that it was an exothermic process and the conformation of aptamer changed during the recognition process. Loop bases in the secondary structure of aptamer formed a special binding pocket to recognize AFB1. Van der Waals and electrostatic interaction were the main driving forces. By blocking the stem bases guided by the structural investigation, a rationally designed CRISPR/Cas12a-Exo III aptasensor for AFB1 detection was constructed, and the sensitivity was improved by target recycling. Under optimal conditions, the linear detection range for AFB1 was 0.01-20 ng/mL, and AFB1 was accurately determined in corn and wheat samples. This work laid a theoretical foundation for the design of AFB1 aptasensor, and the developed detection model came up with new ideas for the development of CRISPR/Cas12a-based aptasensor.
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