2. Academic Validation
  3. DNA Tetrahedron-Enhanced Catalytic Hairpin Assembly for in Situ Monitoring of Viral RNA in Host Cells

DNA Tetrahedron-Enhanced Catalytic Hairpin Assembly for in Situ Monitoring of Viral RNA in Host Cells

  • Chemistry. 2025 Jul 2;31(37):e202501329. doi: 10.1002/chem.202501329.
Sha Lu 1,  Ai-Xin Ma 1,  Xiao-Lin Gao 2,  Zhi-Qi Dai 1,  Qing-Nan Li 1,  Yang-Yang Liu 1,  Yi Zhang 3,  Qi Meng 3,  Dai-Wen Pang 1,  Yun-Xi Cui 3,  De-Ming Kong 1 4
Affiliations

Affiliations

  • 1 State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, P. R. China.
  • 2 School of Agriculture, Food and Ecosystem Sciences, University of Melbourne, Melbourne, Australia.
  • 3 College of Life Sciences, Nankai University, Tianjin, 300071, P. R. China.
  • 4 School of Chemistry and Chemical Engineering, Qinghai Minzu University, Xi'ning, 810008, P. R. China.
PMID: 40407343 DOI: 10.1002/chem.202501329
Abstract

Viruses pose significant threats to global public health and have substantial socio-economic impacts. Developing rapid, sensitive, and specific in situ virus-detection technologies is crucial for tracking viral genome release, replication in host cells, and intercellular transmission, thus holding great promise in understanding viral Infection mechanisms, enabling early diagnosis, screening Antiviral drugs, and formulating prevention and treatment strategies. In this study, we present a multifunctional tetrahedral DNA nanostructure (TDN)-based system designed to monitor viral RNA levels in host cells in situ. The system employs a TDN-based hyperbranched catalytic hairpin assembly (CHA) reaction to achieve rapid and powerful signal amplification, generating specific fluorescence resonance energy transfer (FRET) signals in response to target viral RNA. Using Japanese encephalitis virus (JEV) as a model, we developed the THA@JEV system responding to JEV-RNA. This system demonstrated rapid and sensitive detection of JEV-RNA within 12 minutes in a cell-free environment, enabling real-time imaging of JEV-RNA in situ within host cells. The integrated TDN structure enhanced the biological stability of the probes and ensured their reliability under various stress conditions and microenvironments. This TDN-based sensor system shows significant potential for elucidating viral Infection pathways and advancing related virology research, as well as for screening Antiviral drugs.

Keywords

DNA nanotechnology; DNA tetrahedron; JEV; biosensor; viral infection.

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