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

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

968
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
968
CRISPR01:59

CRISPR

54.7K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
54.7K
Homologous Recombination02:31

Homologous Recombination

58.8K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
58.8K
CRISPR and crRNAs02:53

CRISPR and crRNAs

18.1K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
18.1K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.4K
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...
6.4K

You might also read

Related Articles

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

Sort by
Same author

ESR1 fusion proteins in breast cancer: distinguishing oncogenic drivers from passenger events.

Oncogene·2026
Same author

Reinforced optical cage systems enable drift-free single-molecule localization microscopy.

Communications engineering·2025
Same author

Paclitaxel compromises nuclear integrity in interphase through SUN2-mediated cytoskeletal coupling.

Journal of cell science·2025
Same author

Universal preimplantation genetic testing for monogenic disease (Karyomapping): diagnosis of >1000 unique disorders with no detected misdiagnoses.

Human reproduction (Oxford, England)·2025
Same author

Conserved Phosphorylation of the Myosin1e TH1 Domain Impacts Membrane Association and Function in Yeast and Worms.

Cytoskeleton (Hoboken, N.J.)·2025
Same author

Detecting chromosomal rearrangements in boars using Hi-C.

Animal genetics·2025
Same journal

Correction: Reis et al. Bioinks Enriched with ECM Components Obtained by Supercritical Extraction. <i>Biomolecules</i> 2022, <i>12</i>, 394.

Biomolecules·2026
Same journal

Correction: Kim, K.-H.; Yoo, B.C. Gintonin as a Lysophosphatidic Acid-Enriched GPCR Ligand System: Molecular Architecture and Receptor Pharmacology in <i>Panax ginseng</i>. <i>Biomolecules</i> 2026, <i>16</i>, 465.

Biomolecules·2026
Same journal

Correction: Bastyte et al. The Association of Vitamin D Receptor Gene Polymorphisms with Vitamin D, Total IgE, and Blood Eosinophils in Patients with Atopy. <i>Biomolecules</i> 2024, <i>14</i>, 212.

Biomolecules·2026
Same journal

AtHSPR Plays a Positive Role in Arabidopsis Resistance Against <i>Pseudomonas syringae</i> pv. <i>tomato</i> DC3000 by Interacting with TOP1.

Biomolecules·2026
Same journal

CYTH4 Facilitates Renal Cell Carcinoma via Enhancing Proliferation and Likely Immune Evasion.

Biomolecules·2026
Same journal

Integrated Immune-Gut Profiling Identifies an Exploratory Pediatric Inflammatory Intestinal Profile Associated with Food-Specific IgG Reactivity.

Biomolecules·2026
See all related articles

Related Experiment Video

Updated: Nov 15, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
11:35

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

13.0K

A Targeted and Tuneable DNA Damage Tool Using CRISPR/Cas9.

Ioannis Emmanouilidis1, Natalia Fili2, Alexander W Cook2

  • 1School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK.

Biomolecules
|March 6, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method using Cas9 (CRISPR-associated protein 9) to create precise DNA double-strand breaks (DSBs) in mammalian cells. This controlled DNA damage induction allows for better study of DNA repair pathways at specific genomic locations.

Keywords:
Cas9DNA damageDNA repairdouble-strand break

More Related Videos

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
08:20

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

4.3K
CIRCLE-Seq for Interrogation of Off-Target Gene Editing
08:23

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

1.1K

Related Experiment Videos

Last Updated: Nov 15, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
11:35

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

13.0K
A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
08:20

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

4.3K
CIRCLE-Seq for Interrogation of Off-Target Gene Editing
08:23

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

1.1K

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Mammalian cells possess DNA repair pathways to counteract constant DNA damage.
  • Double-strand breaks (DSBs) are critical DNA lesions impacting cell viability and genome stability.
  • Current methods for inducing DSBs (e.g., irradiation, drugs) lack precision in location and dosage.

Purpose of the Study:

  • To develop a method for inducing targeted and controlled DNA double-strand breaks (DSBs) at specific genomic locations.
  • To enable the study of DNA damage response and repair mechanisms in relation to local chromatin states.
  • To provide a versatile tool for investigating genome stability and repair pathway activation.

Main Methods:

  • Utilized the RNA-guided Cas9 (CRISPR-associated protein 9) endonuclease.
  • Designed custom promiscuous guide RNAs based on in silico predictions.
  • Employed electroporation of recombinant Cas9-guide complexes to induce DSBs in human cell lines.

Main Results:

  • Successfully induced DSBs at defined quantities and specific locations across the human genome.
  • Demonstrated a generic, low-cost, and rapid methodology for controlled DNA damage induction.
  • Established a new model system for studying DNA repair at targeted genomic sites.

Conclusions:

  • The Cas9-mediated induction of targeted DSBs offers a significant advancement over traditional methods.
  • This technique provides unprecedented control for investigating DNA damage response and repair in mammalian cells.
  • The methodology is adaptable for various research applications in genome stability and therapeutic development.