Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

You might also read

Related Articles

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

Sort by
Same author

IRX3 depletion promotes early cardiac commitment of hiPSC-Derived Cardiomyocytes.

PloS one·2026
Same author

EMILIN1 emerges as a TGFβ/SETDB1-regulated secreted biomarker in Duchenne muscular dystrophy.

Cell death & disease·2026
Same author

Functional impact of the eIF6 N106S mutation on ribosome biogenesis in wild type and Shwachman-Diamond syndrome cells.

Molecular medicine (Cambridge, Mass.)·2026
Same author

The hegemonic EWSR1::ETS oncoprotein overrules core regulatory circuitry principles in Ewing sarcoma.

NPJ precision oncology·2025
Same author

Retinal organoids mirror CRISPR-Cas9 gene editing efficiency observed <i>in vivo</i>.

Molecular therapy. Methods & clinical development·2025
Same author

Nuclear hormone-sensitive lipase regulates adipose tissue mass and adipocyte metabolism.

Cell metabolism·2025

Related Experiment Video

Updated: Jul 3, 2026

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
10:07

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

Triplex-forming oligonucleotide-orthophenanthroline conjugates for efficient targeted genome modification.

Fabio Cannata1, Erika Brunet, Loïc Perrouault

  • 1Laboratoire de Biophysique, Centre National de la Recherche Scientifique Unité Mixte de Recherche 5153 USM 503, and Institut National de la Santé et de la Recherche Médicale U565, Muséum National d'Histoire Naturelle, 43 Rue Cuvier, 75005 Paris, France.

Proceedings of the National Academy of Sciences of the United States of America
|July 5, 2008
PubMed
Summary

Synthetic nucleases made from orthophenanthroline (OP) and triplex-forming oligonucleotides (TFOs) efficiently create targeted DNA double-strand breaks (DSBs) for gene modification. This breakthrough enables precise genome editing in cells, paving the way for new therapeutic strategies.

More Related Videos

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
09:04

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids

Published on: September 21, 2017

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks
12:19

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Published on: November 10, 2016

Related Experiment Videos

Last Updated: Jul 3, 2026

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
10:07

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
09:04

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids

Published on: September 21, 2017

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks
12:19

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Published on: November 10, 2016

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Gene modification efficiency is limited by homologous recombination.
  • Creating targeted double-strand breaks (DSBs) is crucial for efficient gene editing.
  • Engineering specific nucleases for targeted gene modification remains a significant challenge.

Purpose of the Study:

  • To develop novel synthetic nucleases for efficient and targeted genome modification.
  • To demonstrate the efficacy of orthophenanthroline-triplex-forming oligonucleotide (OP-TFO) conjugates in inducing targeted DSBs.
  • To assess the potential of OP-TFOs for gene editing applications in cultured cells.

Main Methods:

  • Conjugation of orthophenanthroline (OP), a DNA cleaving molecule, with triplex-forming oligonucleotides (TFOs).
  • Introduction of OP-TFO conjugates into cultured cells to induce targeted DSBs.
  • Analysis of mutations at the target site using nonhomologous end joining (NHEJ) repair.
  • Cotransfection with homologous donor DNA to evaluate targeted gene modification via homologous recombination.

Main Results:

  • OP-TFO conjugates successfully induced targeted DSBs in cultured cells.
  • A specific OP-TFO demonstrated high efficiency, leading to mutations in approximately 10% of treated cells.
  • Targeted gene modification occurred in over 1.5% of cells upon cotransfection with homologous donor DNA.
  • The frequency of triplex-forming sequences in human and mouse genes suggests broad applicability.

Conclusions:

  • OP-TFO conjugates represent a new class of efficient site-specific nucleases for targeted genome modification.
  • This approach overcomes limitations of traditional gene editing methods by harnessing DNA-damaging molecules.
  • The findings open avenues for advancing DNA repair research and cancer studies.