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

65.0K
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...
65.0K
Homologous Recombination02:31

Homologous Recombination

7.2K
7.2K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

7.3K
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...
7.3K
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

2.5K
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...
2.5K
CRISPR01:59

CRISPR

58.8K
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...
58.8K
Gene Conversion02:08

Gene Conversion

10.8K
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
10.8K

You might also read

Related Articles

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

Sort by
Same author

MRP class of human ATP binding cassette (ABC) transporters: historical background and new research directions.

Xenobiotica; the fate of foreign compounds in biological systems·2008
Same author

A biomechanical, radiologic, and clinical comparison of outcome after multilevel cervical laminectomy or laminoplasty in the rabbit.

Spine·2000
Same author

Clinical outcome scales for use in a rabbit model of cervical myelopathy.

Journal of spinal disorders·2000
Same author

The chondroitin sulfate proteoglycans neurocan and phosphacan are expressed by reactive astrocytes in the chronic CNS glial scar.

The Journal of neuroscience : the official journal of the Society for Neuroscience·1999
Same author

Maternal and embryonic expression of zebrafish lef1.

Mechanisms of development·1999
Same author

alyron, an insertional mutation affecting early neural crest development in zebrafish.

Developmental biology·1999

Related Experiment Video

Updated: Mar 17, 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

Precise genome editing by homologous recombination.

K Hoshijima1, M J Jurynec1, D J Grunwald1

  • 1University of Utah, Salt Lake City, UT, United States.

Methods in Cell Biology
|July 23, 2016
PubMed
Summary

This study presents simple and efficient methods for precise zebrafish genome editing using homologous recombination and double-strand breaks. Successfully edited alleles can be identified using an incorporated reporter gene, enhancing gene editing efficiency.

Keywords:
Conditional mutationGenome editingHomologous recombinationKnockin mutationZebrafish

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

2.8K
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

13.4K

Related Experiment Videos

Last Updated: Mar 17, 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

2.8K
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

13.4K

Area of Science:

  • Genetics
  • Molecular Biology
  • Zebrafish Model Organisms

Background:

  • Precise genome modification is crucial for understanding gene function.
  • Zebrafish are a valuable model organism for genetic studies.
  • Existing genome editing techniques can be inefficient or lack precise control.

Purpose of the Study:

  • To develop simple and efficient methods for precise zebrafish genome editing.
  • To enable simultaneous multiple genome modifications at a single locus.
  • To facilitate the identification of successfully edited alleles through reporter gene expression.

Main Methods:

  • Utilizing homologous recombination between host genome and double-stranded DNA (dsDNA) donor molecules.
  • Inducing targeted double-strand breaks to stimulate recombination.
  • Employing I-SceI meganuclease enzyme digestion for effective dsDNA donor molecules.
  • Incorporating linked reporter genes into donor sequences for allele identification.

Main Results:

  • Demonstrated methods for creating simple sequence changes, in-frame additions, and knockin/knockout alleles.
  • Successfully generated conditional alleles with exons flanked by loxP sites.
  • Showcased efficient genome editing in zebrafish with reporter gene-driven identification of edited alleles.

Conclusions:

  • The presented methods offer a robust and efficient approach for precise zebrafish genome editing.
  • Simultaneous modifications and reporter-assisted selection significantly improve genome editing outcomes.
  • These techniques advance the utility of zebrafish as a model for genetic research and disease modeling.