Search research articles

Search research articles

Related Experiment Videos

Protein-free parallel triple-stranded DNA complex formation.

A K Shchyolkina¹, E N Timofeev, Y P Lysov

¹Engelhardt Institute of Molecular Biology, Russian Academy of Science, 117984 Moscow, Russia. annas@genome.eimb.relarn.ru

Nucleic Acids Research

|February 13, 2001

Summary

This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

[Preparation of Modified Combinatorial DNA Libraries via Emulsion PCR with Subsequent Strand Separation].

Molekuliarnaia biologiia·2019

Same author

[Enzymatic synthesis of high-modified DNA].

Molekuliarnaia biologiia·2017

Same author

Elevated α-synuclein caused by SNCA gene triplication impairs neuronal differentiation and maturation in Parkinson's patient-derived induced pluripotent stem cells.

Cell death & disease·2015

Same author

Alpha-Synuclein affects neurite morphology, autophagy, vesicle transport and axonal degeneration in CNS neurons.

Cell death & disease·2015

Same author

Visualization of lipid-receptor interactions on single cells by time-resolved imaging fluorescence microscopy.

Journal of fluorescence·2013

Same author

Targeting duplex DNA with chimeric α,β-triplex-forming oligonucleotides.

Nucleic acids research·2012

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

This study demonstrates that a DNA sequence modified with acridine can form a specific parallel triple helix with a DNA duplex. This finding suggests the possibility of protein-independent parallel triplex formation.

Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

DNA triple helix formation is a key area in molecular biology.
Understanding DNA-protein interactions is crucial for gene regulation.
The ZNF91 gene encodes a human KRAB zinc finger protein.

Purpose of the Study:

To investigate the formation of specific DNA triple helices.
To explore protein-independent triplex formation using modified oligonucleotides.
To confirm the orientation and stability of the formed triplex structures.

Main Methods:

Synthesis of a DNA probe with an acridine intercalator.
Electrophoretic mobility shift assays to determine binding constants.
Fluorescence anisotropy and resonance energy transfer to confirm triplex formation and orientation.

Related Experiment Videos

Chemical modification protection assays (OsO4) to assess strand displacement.

Main Results:

A 14 nt DNA sequence conjugated with acridine formed a specific parallel triple helix with a 16 bp DNA duplex of identical sequence.
Binding was confirmed by electrophoretic band shifts and increased fluorescence anisotropy.
Fluorescence resonance energy transfer confirmed the parallel orientation of the triplex.
Protection from chemical modification indicated no displacement of the target duplex strand.

Conclusions:

Acridine-modified DNA can form stable, protein-independent parallel triple helices.
These findings have implications for understanding DNA structural dynamics and potential therapeutic applications.
The study provides evidence for a novel mode of DNA recognition and binding.