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l-DNA-Based Catalytic Hairpin Assembly Circuit.

Adam M Kabza1, Jonathan T Sczepanski1

  • 1Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.

Molecules (Basel, Switzerland)
|February 26, 2020
PubMed
Summary

We developed a nuclease-resistant DNA circuit using l-DNA for robust signal amplification in biological fluids. This breakthrough enables sensitive detection of disease biomarkers in challenging environments like fetal bovine serum (FBS).

Keywords:
catalytic hairpin assembly (CHA)l-DNAmicroRNApeptide nucleic acidstrand-displacement reaction

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Nanotechnology

Background:

  • Isothermal, enzyme-free DNA amplification shows promise for biosensing and diagnostics.
  • Operating DNA-based systems in biological environments is challenging due to nuclease degradation.

Purpose of the Study:

  • To develop a nuclease-resistant DNA amplification circuit for use in biological environments.
  • To demonstrate the feasibility of detecting nucleic acid biomarkers in complex biological samples.

Main Methods:

  • Constructed a catalytic hairpin assembly (CHA) circuit using nuclease-resistant l-DNA.
  • Compared the biostability of l-DNA CHA circuits with native d-DNA CHA circuits.
  • Interfaced the l-CHA circuit with a d-nucleic acid biomarker using a peptide nucleic acid (PNA) intermediary.

Main Results:

  • The l-DNA CHA circuit exhibited unimpeded signal amplification in 10% fetal bovine serum (FBS).
  • l-DNA CHA demonstrated superior biostability compared to d-DNA CHA.
  • Sequence-specific detection of an endogenous d-nucleic acid biomarker in FBS was achieved.

Conclusions:

  • l-DNA CHA circuits provide a robust platform for nucleic acid detection in harsh biological settings.
  • This approach enables the detection of low-abundance nucleic acids.
  • The study supports the use of l-oligonucleotides in DNA nanotechnology for diagnostics.