An Ultra-Stable Threose Nucleic Acid-Based Biosensors for Rapid and Sensitive Nucleic Acid Detection and in Vivo Imaging

Abstract

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 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 GO-TNA. It comprises chemically modified TNA capture probes on graphene oxide (GO) for detecting and imaging target nucleic acids in vitro and in vivo, distinguishing single nucleobase mismatches, and monitoring dynamic changes in target miRNA. By loading TNA capture probes onto the GO substrate, the GO-TNA sensing platform for nucleic acid detection demonstrates a significant 1000-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. Additionally, it has potential applications in environmental analysis and food safety, utilizing different TNA-based capture probe sequences to detect pesticides, antibiotics, and foodborne pathogens.

Note:

Funding Information: This work is supported by Health and Medical Research Fund (07181396 and 05160336), National Science Foundation of China (21574109 and 217780430), Hong Kong Research Grants Council 11307421, 11301220, 11304719, The Science and Technology Innovation Committee of Shenzhen Municipality (JCYJ20190812160203619), City University of Hong Kong 7005832, 9680104 and 7004911, and Dongguan Science and Technology of Social Development Program (20231800912372 and 20231800925372).

Declaration of Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Ethics Approval Statement: All animal experiments were approved by the Animal Welfare and Ethics Committee, and conducted in accordance with the Guidelines for Institutional Animal Care and Use of Affiliated Dongguan Hospital, Southern Medical University

Keywords: Unnatural TNAs, Capture probes, Biosensors, Graphene oxide, Nucleic acid detection