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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...
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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...
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Updated: Jul 11, 2026

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
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Published on: January 8, 2015

Road to precision: recombinase-based targeting technologies for genome engineering.

Dagmar Wirth1, Leonor Gama-Norton, Pamela Riemer

  • 1Helmholtz Centre for Infection Research, Department of Gene Regulation and Differentiation and Model Systems for Infection and Immunity, Inhoffenstrasse 7, D-38124 Braunschweig, Germany. Dagmar.Wirth@helmholtz-hzi.de

Current Opinion in Biotechnology
|October 2, 2007
PubMed
Summary

Recombinase-mediated cassette exchange (RMCE) precisely integrates transgenes into mammalian cells. This review details targeting strategies using Cre, Flp, and PhiC31 recombinases for applications like protein expression and transgenic animal creation.

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Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
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Published on: January 8, 2015

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Area of Science:

  • Genetics
  • Molecular Biology
  • Mammalian Cell Engineering

Background:

  • Recombinase-based gene targeting offers precise transgene integration into mammalian genomes.
  • This precision allows for predictable expression of introduced genes.
  • Site-specific recombinases are key tools in modern genetic engineering.

Purpose of the Study:

  • To review current recombinase-based strategies for transgene integration.
  • To highlight the applications of these targeting methods.
  • To discuss the use of Cre, Flp, and PhiC31 recombinases.

Main Methods:

  • Review of literature on recombinase-mediated cassette exchange (RMCE).
  • Focus on site-specific recombinases like Cre, Flp, and PhiC31.
  • Description of targeting strategies and their applications.

Main Results:

  • RMCE enables precise integration of transgenes into specific chromosomal locations.
  • Recombinase systems facilitate predictable gene expression in modified cells.
  • Various applications including protein production, viral vector development, and creating transgenic organisms are enabled.

Conclusions:

  • Recombinase-based targeting is a powerful approach for mammalian cell engineering.
  • These methods offer significant advantages for predicting transgene expression.
  • Applications span diverse fields from basic research to biotechnology.