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Related Experiment Video

Updated: Nov 14, 2025

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique
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Oriented Soft DNA Curtains for Single-Molecule Imaging.

Aurimas Kopu Stas1, Šaru Nė Ivanovaitė, Tomas Rakickas

  • 1Life Sciences Center, Institute of Biotechnology, Vilnius University, Saulėtekio av. 7, LT-10257 Vilnius, Lithuania.

Langmuir : the ACS Journal of Surfaces and Colloids
|March 10, 2021
PubMed
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New DNA Curtains use protein templates for stable, oriented DNA stretching, enabling precise study of protein-DNA interactions like CRISPR-Cas9 binding. This advancement enhances biophysical studies with faster data acquisition and controlled conditions.

Area of Science:

  • Biophysics
  • Nanotechnology
  • Molecular Biology

Background:

  • Single-molecule methods are crucial for biophysical studies.
  • Nanotechnology platforms enhance experimental design and data acquisition speed.
  • DNA Curtains are a key nanotechnological platform for DNA stretching.

Purpose of the Study:

  • To develop advanced strategies for fabricating stable, oriented DNA molecule arrays using protein template-directed assembly.
  • To improve DNA molecule immobilization and extension for controlled biophysical experiments.
  • To demonstrate the utility of the improved DNA Curtains for studying protein-DNA interactions.

Main Methods:

  • Fabrication of protein templates on glass coverslips for directional DNA assembly.
  • Utilizing biotinylated DNA molecules for immobilization onto protein templates.

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Last Updated: Nov 14, 2025

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Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA
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  • Employing single-end and both-end immobilization strategies, including heterologous DNA labeling and antibody use.
  • Using traptavidin for enhanced immobilization stability.
  • Demonstrating the assay with fluorescently labeled nucleic acid-interacting proteins, such as CRISPR-Cas9.

    • Main Results:

    • Successfully created oriented arrays of stably immobilized DNA molecules using protein templates.
    • Developed a both-end immobilization method that eliminates the need for continuous buffer flow, allowing for more controlled reaction conditions.
    • Achieved enhanced immobilization stability of DNA molecules.
    • Validated the double-tethered Soft DNA Curtains for monitoring protein binding location and position on individual DNA molecules.

    Conclusions:

    • The developed protein template-directed assembly offers a robust method for creating stable, oriented DNA Curtains.
    • Both-end immobilization provides superior control over reaction conditions for studying protein-DNA interactions.
    • This improved DNA Curtains platform is effective for high-resolution analysis of nucleic acid-binding proteins, including CRISPR-Cas9.