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

65.3K
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...
65.3K
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

2.6K
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...
2.6K
CRISPR01:59

CRISPR

58.9K
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...
58.9K
CRISPR01:59

CRISPR

18.9K
18.9K
CRISPR and crRNAs02:53

CRISPR and crRNAs

19.5K
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...
19.5K

You might also read

Related Articles

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

Sort by
Same author

Author Correction: Forecasting the dissemination of antibiotic resistance genes across bacterial genomes.

Nature communications·2026
Same author

Reply to: Genome contamination may lead to an overestimation of horizontal gene transfer inferences.

Nature communications·2026
Same author

Metabolic, enterohepatic and gut microbial effects of atorvastatin in healthy men.

Endocrine connections·2026
Same author

Analysis of Proteome Dynamics in Early-Stage Sporulation-Inhibited Variants of <i>Parageobacillus thermoglucosidasius</i>.

International journal of molecular sciences·2025
Same author

Gut microbiome-mediated transformation of dietary phytonutrients is associated with health outcomes.

Nature microbiology·2025
Same author

Fecal exfoliome sequencing captures immune dynamics of the healthy and inflamed gut.

Nature biotechnology·2025
Same journal

Correction: A method for supervoxel-wise association studies of age and other non-imaging variables from coronary computed tomography angiograms.

Scientific reports·2026
Same journal

Poly(bromophenol blue)/CoSn(OH)<sub>6</sub> cubic particles modified pencil graphite electrode for electrochemical determination of diphenhydramine.

Scientific reports·2026
Same journal

Dietary Chlorella, Spirulina, and acidifier modulate jejunal cytokine-related gene expression in broiler chickens.

Scientific reports·2026
Same journal

Perceived physical activity barriers in university students: associations with fatigue and eating behaviours.

Scientific reports·2026
Same journal

Refuge limitation structures habitat use in agricultural landscapes: evidence from Sunda pangolins.

Scientific reports·2026
Same journal

Lightweight stateless transaction verification with outsourced witness updates for UTXO blockchains.

Scientific reports·2026
See all related articles

Related Experiment Video

Updated: Mar 26, 2026

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

17.2K

CRMAGE: CRISPR Optimized MAGE Recombineering.

Carlotta Ronda1, Lasse Ebdrup Pedersen1, Morten O A Sommer1

  • 1The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kogle Allé 6, 2970 Hørsholm, Denmark.

Scientific Reports
|January 23, 2016
PubMed
Summary
This summary is machine-generated.

We developed CRMAGE, a CRISPR/Cas9 and MAGE-based genome engineering tool for Escherichia coli. This method significantly enhances recombineering efficiency for faster and more precise genetic modifications.

More Related Videos

Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice
06:46

Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice

Published on: April 2, 2020

10.6K
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

Related Experiment Videos

Last Updated: Mar 26, 2026

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

17.2K
Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice
06:46

Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice

Published on: April 2, 2020

10.6K
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

Area of Science:

  • Microbiology
  • Molecular Biology
  • Synthetic Biology

Background:

  • Efficient genome engineering is crucial for metabolic engineering and systems biology.
  • Current genome engineering technologies for bacteria like Escherichia coli have limitations in efficiency and speed.

Purpose of the Study:

  • To develop a highly efficient and rapid genome engineering method for Escherichia coli.
  • To overcome the bottleneck of low efficiency in traditional recombineering techniques.

Main Methods:

  • Combined CRISPR/Cas9 and lambda Red recombineering-based MAGE technology (CRMAGE).
  • Utilized USER-cloning for plasmid assembly and CRISPR/Cas9 for plasmid curing.
  • Developed a web-based tool for designing guide RNAs (gRNAs) and oligos.

Main Results:

  • Achieved 96.5-99.7% recombineering efficiency for gene recoding, a significant improvement over traditional methods (0.68-5.4%).
  • Increased efficiency for protein synthesis modulation (insertions/RBS substitutions) from 6% to 70%.
  • Demonstrated multiplexing capability, allowing introduction of multiple mutations in one round, and targeted PAM-independent loci.

Conclusions:

  • CRMAGE offers a substantial advancement in genome engineering efficiency and speed for Escherichia coli.
  • The platform facilitates multiple engineering rounds per day and holds potential for automating genome-scale engineering.