HAT probe with adjustable switching characteristic and application thereof

By designing the polydeoxythymidine sequence and raised secondary structure of the HAT probe, combined with the DNAzyme activation mechanism and AuNP signal enrichment, the problems of false positives and signal leakage of nucleic acid probes in complex samples were solved, and high sensitivity and stable biosensing effect were achieved.

CN122104684APending Publication Date: 2026-05-29TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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Patent Information

Application Number
CN202610071204.7
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-01-20
Publication Date
2026-05-29

AI Technical Summary

Technical Problem

Existing nucleic acid probes are prone to non-specific binding in complex biological samples, leading to false positives. Furthermore, the signal amplification process is prone to leakage, resulting in poor system robustness and a lack of external control capabilities. It is difficult to achieve high sensitivity and low background signal without affecting the target binding ability.

Method used

A HAT probe containing a polydeoxythymidine sequence and a raised secondary structure was designed to inhibit Cas12a trans-cleavage through steric hindrance, and to achieve fine regulation of Cas12a activity by combining DNAzyme activation mechanism and AuNP signal enrichment.

Benefits of technology

It achieves high signal-to-noise ratio biosensing, maintains high sensitivity and stability in complex samples, has a detection limit of up to 2.05×10-15M, and features low background and fast response capabilities.

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Abstract

The invention discloses an HAT probe with controllable switching characteristics and application thereof, the HAT probe comprises a secondary structure capable of forming specific steric hindrance, the secondary structure is a raised bubble or ring, the core of the secondary structure is a polydeoxythymidine sequence, and the length of the polydeoxythymidine sequence is between 5-30 nucleotides. The HAT probe disclosed by the invention has a'switch 'regulation function; and a molecular switch is constructed by utilizing the intact structure and the cut linear structure of the HAT probe to inhibit and activate the activity of Cas12a. The'switch 'is capable of responding to specific biomolecules (such as miRNA and protein) and converting the existence of the biomolecules into a detectable signal.
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