Department of Chemistry and State Key Laboratory of Marine Pollution, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong, Hong Kong SAR, P. R. China.
Shenzhen Clinical Medical College, Guangzhou University of Chinese Medicine, 518116 Shenzhen, P. R. China.
Acta Biomater. 2024 Mar 15;177:472-485. doi: 10.1016/j.actbio.2024.01.031. Epub 2024 Feb 1.
The human genome's nucleotide sequence variation, such as single nucleotide mutations, can cause numerous genetic diseases. However, detecting nucleic acids accurately and rapidly in complex biological samples remains a major challenge. While natural deoxyribonucleic acid (DNA) has been used as biorecognition probes, it has limitations like poor specificity, reproducibility, nuclease-induced enzymatic degradation, and reduced bioactivity on solid surfaces. To address these issues, we introduce a stable and reliable biosensor called graphene oxide (GO)- threose nucleic acid (TNA). It comprises chemically modified TNA capture probes on GO for detecting and imaging target nucleic acids in vitro and in vivo, distinguishing single nucleobase mismatches, and monitoring dynamic changes in target microRNA (miRNA). By loading TNA capture probes onto the GO substrate, the GO-TNA sensing platform for nucleic acid detection demonstrates a significant 88-fold improvement in the detection limit compared to TNA probes alone. This platform offers a straightforward preparation method without the need for costly and labor-intensive isolation procedures or complex chemical reactions, enabling real-time analysis. The stable TNA-based GO sensing nanoplatform holds promise for disease diagnosis, enabling rapid and accurate detection and imaging of various disease-related nucleic acid molecules at the in vivo level. STATEMENT OF SIGNIFICANCE: The study's significance lies in the development of the GO-TNA biosensor, which addresses limitations in nucleic acid detection. By utilizing chemically modified nucleic acid analogues, the biosensor offers improved reliability and specificity, distinguishing single nucleobase mismatches and avoiding false signals. Additionally, its ability to detect and image target nucleic acids in vivo facilitates studying disease mechanisms. The simplified preparation process enhances practicality and accessibility, enabling real-time analysis. The biosensor's potential applications extend beyond healthcare, contributing to environmental analysis and food safety. Overall, this study's findings have substantial implications for disease diagnosis, biomedical research, and diverse applications, advancing nucleic acid detection and its impact on various fields.
人类基因组的核苷酸序列变异，如单核苷酸突变，可导致多种遗传疾病。然而，在复杂的生物样本中准确快速地检测核酸仍然是一个主要挑战。虽然天然脱氧核糖核酸（DNA）已被用作生物识别探针，但它存在特异性差、重现性差、核酸酶诱导的酶降解以及在固体表面上生物活性降低等局限性。为了解决这些问题，我们引入了一种称为氧化石墨烯（GO）- threose 核酸（TNA）的稳定可靠的生物传感器。它由 GO 上的化学修饰 TNA 捕获探针组成，用于体外和体内检测和成像靶核酸，区分单核苷酸错配，并监测靶 microRNA（miRNA）的动态变化。通过将 TNA 捕获探针加载到 GO 基底上，GO-TNA 用于核酸检测的传感平台与单独的 TNA 探针相比，检测限提高了 88 倍。该平台提供了一种简单的制备方法，无需昂贵且劳动密集型的分离程序或复杂的化学反应，实现了实时分析。基于 TNA 的稳定 GO 传感纳米平台有望用于疾病诊断，能够快速准确地检测和成像各种与疾病相关的核酸分子在体内水平。
这项研究的意义在于开发 GO-TNA 生物传感器，该传感器解决了核酸检测的局限性。通过利用化学修饰的核酸类似物，该生物传感器提供了更高的可靠性和特异性，区分单核苷酸错配并避免假信号。此外，它在体内检测和成像靶核酸的能力有助于研究疾病机制。简化的制备过程增强了实用性和可及性，实现了实时分析。生物传感器的潜在应用不仅限于医疗保健，还涉及环境分析和食品安全。总的来说，这项研究的发现对疾病诊断、生物医学研究和各种应用具有重要意义，推动了核酸检测及其对各个领域的影响。
