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Related Concept Videos

Amplifying Signals via Enzymatic Cascade01:22

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When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze...
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Author Spotlight: Advancements in DNA Nanosensors – Addressing Sensitivity and Selectivity Challenges in Molecular Detection
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Sensitive Detection of Nucleic Acids Using Subzyme Feedback Cascades.

Nicole Hasick1,2, Andrea Lawrence1, Radhika Ramadas1

  • 1SpeeDx, Pty Ltd., Eveleigh, NSW 2015, Australia.

Molecules (Basel, Switzerland)
|April 16, 2020
PubMed
Summary

A new DNAzyme feedback system enhances detection sensitivity for nucleic acid biomarkers. This cross-catalytic cascade amplifies signals, enabling rapid and affordable detection of low target concentrations.

Keywords:
DNAzymePlexZymecatalytic DNAdeoxyribozymefeedback cascadeisothermalpoint of caresignal amplification

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Subzymes offer controlled DNAzyme activity for nucleic acid detection.
  • Subzymes alone have limited sensitivity for low target concentrations.

Purpose of the Study:

  • To develop a highly sensitive detection method for nucleic acids.
  • To improve upon existing Subzyme sensitivity using a feedback system.

Main Methods:

  • A cross-catalytic Subzyme feedback system was designed using microparticle-tethered DNAzymes separated by a membrane.
  • Target-induced cleavage initiates a cascade, releasing DNAzymes to interact and amplify the signal.
  • Activated DNAzymes cleave fluorescent substrates for signal generation.

Main Results:

  • The system detected 1 femtomolar (fM) DNA of Chlamydia trachomatis ompA gene in 30 minutes.
  • Achieved a 10,000-fold increase in sensitivity compared to DNAzyme detection alone.
  • Demonstrated a universal, isothermal, and protein-enzyme-free detection method.

Conclusions:

  • The Subzyme cascade significantly enhances sensitivity for nucleic acid detection.
  • This method holds potential for rapid, affordable biomarker detection.
  • The universal design allows for adaptation to various target molecules.