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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.
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Related Experiment Video

Updated: Jul 9, 2026

Engineering Cell-permeable Protein
21:08

Engineering Cell-permeable Protein

Published on: December 28, 2009

Stem cell engineering using transducible Cre recombinase.

Lars Nolden1, Frank Edenhofer, Michael Peitz

  • 1Institute of Reconstructive Neurobiology, University of Bonn-Life & Brain Center and Hertie Foundation, Germany.

Methods in Molecular Medicine
|December 19, 2007
PubMed
Summary

Efficient genetic engineering of embryonic stem cells (ES cells) is crucial. This study introduces a novel method using protein transduction of Cre recombinase for rapid, highly efficient conditional mutagenesis in ES cells and their progeny.

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Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange
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Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange

Published on: April 27, 2017

Area of Science:

  • Stem cell biology
  • Genetics
  • Molecular biology

Background:

  • Embryonic stem (ES) cells are vital for regenerative medicine and studying tissue development.
  • Efficient genetic engineering is needed for ES cell applications, including disease modeling and lineage specification.
  • Current methods using Cre recombinase have limitations like random integration and leaky expression.

Purpose of the Study:

  • To develop a rapid and highly efficient method for conditional mutagenesis in ES cells.
  • To overcome limitations of existing Cre-based genetic engineering techniques.
  • To enable precise genetic modifications in ES cells and their differentiated progeny.

Main Methods:

  • Utilizing the Cre/loxP recombination system combined with direct intracellular delivery of Cre protein via protein transduction.
  • Generating purified, transducible Cre protein from Escherichia coli.
  • Introducing Cre protein into human ES (hES) and mouse ES (mES) cells and their neural progeny.

Main Results:

  • Achieved virtually 100% recombination efficiency in both hES and mES cells.
  • Demonstrated rapid and highly efficient conditional mutagenesis.
  • Successfully transduced Cre protein into ES cells and their neural progeny.

Conclusions:

  • Protein transduction of Cre recombinase offers a powerful and efficient strategy for genetic engineering of ES cells.
  • This method overcomes key limitations of traditional Cre-mediated genetic modification.
  • Enables precise manipulation of the ES cell genome for research and therapeutic applications.