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

Homologous Recombination02:31

Homologous Recombination

64.4K
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...
64.4K
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

2.2K
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...
2.2K
CRISPR01:59

CRISPR

58.3K
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...
58.3K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

7.0K
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...
7.0K
CRISPR and crRNAs02:53

CRISPR and crRNAs

19.4K
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...
19.4K

You might also read

Related Articles

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

Sort by
Same author

Moisture-Gated Synergistic Rapid Crystal-to-Liquid Transition in Pyridinium Halide Crystals via [2 + 2] Photocycloaddition.

Small (Weinheim an der Bergstrasse, Germany)·2026
Same author

A healthy lifestyle is associated with lower risk of depression in type 2 diabetes, irrespective of genetic susceptibility: A UK biobank cohort study.

Journal of affective disorders·2026
Same author

Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis.

Nature communications·2026
Same author

Thermally Facilitated Visible-Light-Induced Crystal Melting Stimulated by Photoisomerization for Rapid and Reversible Adhesion.

ACS applied materials & interfaces·2026
Same author

[Cloning, expression pattern, and functional verification of phosphate transporter protein SmPht5;1 in Salvia miltiorrhiza].

Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica·2026
Same author

Fabricating Inorganic/Organic S-Scheme Heterojunction for Efficient Photocatalytic Production of H<sub>2</sub> and H<sub>2</sub>O<sub>2</sub>.

Research (Washington, D.C.)·2026

Related Experiment Video

Updated: Mar 2, 2026

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
08:22

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Published on: March 12, 2018

15.7K

Homology-mediated end joining-based targeted integration using CRISPR/Cas9.

Xuan Yao1,2, Xing Wang1,2, Xinde Hu1

  • 1Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.

Cell Research
|May 20, 2017
PubMed
Summary
This summary is machine-generated.

A new homology-mediated end joining (HMEJ) strategy enhances transgene integration efficiency. This CRISPR/Cas9-based method shows promise for gene editing in animal models and gene therapies.

More Related Videos

Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology
10:07

Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology

Published on: March 16, 2022

2.7K
Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
06:10

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates

Published on: May 9, 2025

1.0K

Related Experiment Videos

Last Updated: Mar 2, 2026

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
08:22

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Published on: March 12, 2018

15.7K
Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology
10:07

Construction of Homozygous Mutants of Migratory Locust Using CRISPR/Cas9 Technology

Published on: March 16, 2022

2.7K
Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
06:10

Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates

Published on: May 9, 2025

1.0K

Area of Science:

  • Molecular Biology
  • Gene Editing
  • Genomics

Background:

  • Targeted transgene integration is crucial for gene editing applications.
  • Current methods like homologous recombination (HR) have limitations in efficiency, especially in non-dividing cells and embryos.
  • Alternative strategies like microhomology-mediated end joining (MMEJ) and non-homologous end joining (NHEJ) offer some improvements but are not universally efficient.

Purpose of the Study:

  • To develop and evaluate a novel homology-mediated end joining (HMEJ) strategy for efficient transgene integration.
  • To compare the efficacy of the HMEJ strategy against established methods (HR, MMEJ, NHEJ) across various cell types and in vivo models.

Main Methods:

  • Utilized CRISPR/Cas9 to induce targeted DNA cleavage in both a transgene donor vector and the genome.
  • The donor vector contained guide RNA target sites and homology arms for HMEJ.
  • Tested the HMEJ strategy in mouse embryonic stem cells, N2a neuroblastoma cells, HEK293T cells, primary astrocytes, neurons, and in vivo models (mouse and monkey embryos, mouse hepatocytes, and neurons).

Main Results:

  • HMEJ did not significantly improve targeting efficiency in mouse embryonic stem cells or N2a cells compared to HR.
  • HMEJ demonstrated higher knock-in efficiency in HEK293T cells, primary astrocytes, and neurons.
  • Significantly enhanced transgene integration efficiency was observed in mouse and monkey embryos, as well as in vivo hepatocytes and neurons, surpassing HR, NHEJ, and MMEJ strategies.

Conclusions:

  • The HMEJ-based strategy offers a more efficient approach for transgene integration compared to HR, NHEJ, and MMEJ in various cell types and in vivo.
  • This method holds significant potential for applications in generating animal models through gene editing and for advancing targeted gene therapies.