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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

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...
Viral Recombination00:57

Viral Recombination

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.
Homologous Recombination02:31

Homologous Recombination

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...
Homologous Recombination02:31

Homologous Recombination

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...
Recombinant DNA01:09

Recombinant DNA

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Related Experiment Video

Updated: Jun 5, 2026

Site-specific Bacterial Chromosome Engineering: &#934;C31 Integrase Mediated Cassette Exchange (IMCE)
08:21

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)

Published on: March 16, 2012

PhiC31 integrase facilitates genetic approaches combining multiple recombinases.

Claudio Monetti1, Koichiro Nishino, Steffen Biechele

  • 1Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Canada.

Methods (San Diego, Calif.)
|December 28, 2010
PubMed
Summary

Scientists developed a new method for genetic engineering using PhiC31 integrase and FLP recombinase in mouse embryonic stem (ES) cells. This approach enables precise DNA insertion and replacement, preserving other recombinases for future genetic modifications.

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Published on: February 2, 2021

Area of Science:

  • Molecular Biology
  • Genetics
  • Genetic Engineering

Background:

  • Homologous and site-specific DNA recombination are crucial for genetic engineering.
  • Cre and FLP recombinases are widely used but leave residual sites, limiting further manipulation.
  • Additional recombinases are needed to expand genome manipulation possibilities in ES cells.

Purpose of the Study:

  • To develop a novel method for site-specific DNA insertion and replacement in mouse ES cells.
  • To combine PhiC31 integrase and FLP recombinase for enhanced genome engineering.
  • To create versatile "dockable" ES cell lines for future genetic modifications.

Main Methods:

  • Utilized PhiC31 integrase and FLP recombinase for site-specific DNA integration.
  • Employed pre-inserted docking sites in mouse ES cell genomes.
  • Implemented a selection strategy using a silent marker activated by a plasmid-delivered promoter.

Main Results:

  • Successfully achieved site-specific insertion and replacement of DNA sequences in mouse ES cells.
  • Demonstrated the ability to integrate any sequence of interest into a predefined locus.
  • Enabled the availability of Cre recombinase for subsequent genetic applications.

Conclusions:

  • The developed method provides a reliable system for creating "dockable" mouse ES cell lines.
  • This technology expands the possibilities for complex genome manipulation and genetic engineering.
  • The approach reduces the need for extensive molecular screening, enhancing efficiency.