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

80
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...
80
CRISPR01:59

CRISPR

52.5K
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...
52.5K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.1K
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...
17.1K
Homologous Recombination02:31

Homologous Recombination

50.7K
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...
50.7K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
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...
6.0K

You might also read

Related Articles

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

Sort by
Same author

Human neonatal MR1T cells have more diverse TCR repertoires but reduced bacterial recognition than adult MR1T cells.

Nature communications·2026
Same author

Generation of a rhesus macaque harboring a multivalent reporter for assessing gene editing outcomes.

Scientific reports·2026
Same author

Distinct phases of immune system programming during ART-suppressed immunodeficiency virus infection.

bioRxiv : the preprint server for biology·2026
Same author

Systemic Interleukin-4 Application Promotes Functional Recovery and Reprograms Neuroinflammatory and Molecular Responses after Spinal Cord Injury in Rats.

Theranostics·2026
Same author

Analysis of Cytosine Base Editors in Bovine Zygotes: Efficiency and Editing Window Characterization Through Targeting the <i>MYO7A</i> Gene.

Current issues in molecular biology·2026
Same author

BCG Vaccination at Birth Shapes the TCR Usage and Functional Profile of MR1T Cells at 9 Weeks of Age.

bioRxiv : the preprint server for biology·2025

Related Experiment Video

Updated: Jul 30, 2025

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System
14:46

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System

Published on: May 28, 2015

11.1K

A Multifunctional and Highly Adaptable Reporter System for CRISPR/Cas Editing.

Jochen M Wettengel1,2, Lea Hansen-Palmus2, Sofiya Yusova1

  • 1Vaccine & Gene Therapy Institute, Oregon Health & Science University, Beaverton, OR 97006, USA.

International Journal of Molecular Sciences
|May 13, 2023
PubMed
Summary

We developed BETLE, a novel reporter system for CRISPR/Cas genome editing. This tool enables fast, sensitive, and cell-specific detection of gene editing and homology-directed repair (HDR) for optimized therapeutic applications.

Keywords:
CRISPR-Cas reporter systemCas9 activityHDRNHEJasCas12asaCas9spCas9

More Related Videos

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

34.1K
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

4.2K

Related Experiment Videos

Last Updated: Jul 30, 2025

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System
14:46

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System

Published on: May 28, 2015

11.1K
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

34.1K
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

4.2K

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • CRISPR/Cas systems offer powerful therapeutic genome editing capabilities.
  • Optimal design of CRISPR guides and homology-directed repair (HDR) templates is crucial for effective genome editing.
  • Current validation and optimization rely on reporter systems, necessitating improved tools.

Purpose of the Study:

  • To introduce BETLE, a novel reporter system for rapid, sensitive, and cell-specific detection of genome editing and HDR.
  • To enable validation and optimization of CRISPR/Cas systems and HDR templates.
  • To facilitate the development of advanced genome editing tools for therapeutic applications.

Main Methods:

  • Developed BETLE, a reporter system encoding multiple reporter proteins in distinct open-reading frames.
  • Utilized out-of-frame non-homologous end joining (NHEJ) to express NanoLuc luciferase for sensitive editing analysis or mTagBFP2 for cell localization.
  • Integrated a validation site for CRISPR/Cas systems and evaluated BETLE with spCas9, saCas9, and asCas12a using a defective moxGFP reporter.

Main Results:

  • BETLE enables fast, sensitive, and cell-specific detection of genome editing and HDR.
  • The system allows for low-cost analysis via NanoLuc luciferase and enumeration/localization via mTagBFP2.
  • Demonstrated successful editing and HDR validation with various CRISPR/Cas nucleases and reporter configurations.

Conclusions:

  • BETLE is a versatile and adaptable tool for rapid detection and optimization of CRISPR/Cas genome editing and HDR.
  • The reporter system facilitates efficient validation of CRISPR/Cas nucleases and HDR templates.
  • BETLE represents a state-of-the-art technology for advancing in vitro and in vivo genome editing applications.