Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

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...

DNA-only Transposons

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...

CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

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...

You might also read

Related Articles

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

Sort by

Same author

Novel <i>Escherichia coli Dhillonvirus</i> DH23 is Able to Grow on the Host Cells at Late Growth Phases.

PHAGE (New Rochelle, N.Y.)·2026

Same author

A Forgotten Phage Thinker: Bacteriophage Research in USSR in 1930s and the Contribution by Professor M.I. Lurie.

PHAGE (New Rochelle, N.Y.)·2026

Same author

Reply to: About proteins of a siphophage tail tip complex reverting to their pre-ejection fold after DNA ejection.

Nature communications·2025

Same author

Isolation and complete genome sequence of <i>Aeromonas</i> bacteriophage Gekk3-15.

F1000Research·2024

Same author

Bacterial Virus Forcing of Bacterial O-Antigen Shields: Lessons from Coliphages.

International journal of molecular sciences·2023

Same author

Nearly complete structure of bacteriophage DT57C reveals architecture of head-to-tail interface and lateral tail fibers.

Nature communications·2023

Search research articles

Related Experiment Video

Updated: Aug 19, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

New Effective Method of Lactococcus Genome Editing Using Guide RNA-Directed Transposition.

Pavel Yu Pechenov¹, Danil A Garagulya¹, Daniil S Stanovov¹

¹Federal Research Center "Fundamentals of Biotechnology" of Russian Academy of Sciences, Leninsky Prospect, 33, Build. 2, 119071 Moscow, Russia.

International Journal of Molecular Sciences

|November 26, 2022

Summary

Researchers improved genome editing in Lactococcus lactis, a key industrial bacterium. The enhanced CRISPR-Cas system efficiently incorporates large DNA fragments up to 10 kbp into the bacterial chromosome.

Keywords:

Lactococcus lactis genomic engineering transposon-associated CRISPR–Cas system

More Related Videos

CRISPR Guide RNA Cloning for Mammalian Systems

CRISPR Guide RNA Cloning for Mammalian Systems

Published on: October 2, 2018

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

Published on: September 19, 2018

Related Experiment Videos

Last Updated: Aug 19, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

CRISPR Guide RNA Cloning for Mammalian Systems

CRISPR Guide RNA Cloning for Mammalian Systems

Published on: October 2, 2018

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

An Efficient Strategy for Generating Tissue-specific Binary Transcription Systems in Drosophila by Genome Editing

Published on: September 19, 2018

Area of Science:

Microbiology
Molecular Biology
Biotechnology

Background:

Lactococcus lactis is a crucial industrial microorganism and model organism for lactic acid bacteria (LAB) research.
Current genome-editing tools limit the incorporation of large genetic materials into LAB chromosomes, hindering strain development.
Developing improved LAB strains for applications like phage resistance and vaccines requires efficient genome modification.

Purpose of the Study:

To enhance genome editing tools for Lactococcus lactis.
To enable the stable incorporation of large DNA fragments (up to 10 kbp) into the L. lactis chromosome.
To facilitate the development of novel LAB strains with improved industrial or research properties.

Main Methods:

Utilized a redesigned CRISPR-Cas-associated transposon system for genome editing in L. lactis.
Optimized experimental conditions to improve the efficiency and stability of genetic material incorporation.
Targeted the L. lactis beta-galactosidase gene for insertion of novel genetic fragments.

Main Results:

Successfully achieved stable chromosomal integration of DNA fragments up to 10 kbp in L. lactis.
Demonstrated editing efficiencies of 2 × 10^-4 for 1 kbp and 4 × 10^-5 for 10 kbp fragments under optimized conditions.
The system proved effective for incorporating significant amounts of novel genetic material.

Conclusions:

The redesigned CRISPR-Cas-associated transposon system offers a powerful tool for efficient genome editing in L. lactis.
This advancement overcomes previous limitations in incorporating large genetic elements, paving the way for improved LAB strain development.
The tool is suitable for fast and straightforward genetic modifications, particularly when coupled with positive selection markers.