Enzymatic Cycle-Inspired Dynamic Biosensors Affording No False-Positive Identification

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Biosensor Technology

Background:

• Accurate nucleic acid detection in clinical samples is crucial for diagnosis.
• Current methods face challenges with false positives from nonspecific interactions.
• Distinguishing true from false positives remains a key challenge in nucleic acid screening.

Purpose of the Study:

• To develop advanced biosensors for reliable nucleic acid detection.
• To overcome limitations of current methods by minimizing false-positive identifications.
• To create dynamic biosensors capable of repeated measurements for enhanced accuracy.

Main Methods:

• Developed an allosteric tool with invertible cooperativity for biosensing probe control.
• Utilized toehold displacement and enzymatic reactions for effector silencing and regeneration.
• Engineered dynamic biosensors for repeated measurement of single nucleic acid samples.

Main Results:

• Demonstrated reversible or cyclic activity control of biosensing probes via allosteric effectors.
• Showcased the ability of dynamic biosensors to repeatedly measure a single nucleic acid sample.
• Established that signal variation diversity distinguishes specific target binding from nonspecific interactions.

Conclusions:

• Dynamic biosensors precisely identify true-positive nucleic acid signals by analyzing diverse signal variations across repeated measurements.
• Nonspecific interactions yield equivalent signal variations, allowing for effective false-positive discrimination.
• These novel biosensors present significant opportunities for improved physiological and pathological diagnosis.