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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...
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...

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Updated: May 13, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

Engineering a Membrane-Anchored CreER Recombinase With Reduced Basal Activity for In Vitro Recombination.

Xiaotong Zeng1, Xinyi Chen1, Zishan Liang1

  • 1Department of Biology, Shantou University, Shantou, Guangdong, China.

Biotechnology Journal
|May 12, 2026
PubMed
Summary
This summary is machine-generated.

A new membrane-tethered CreER-loxP (mCreER-loxP) system reduces unintended cell labeling in genetic studies. This engineered system improves temporal and spatial precision for cell lineage tracing and genetic engineering applications.

Keywords:
Cre‐loxP systemcell labelingmembrane tetheringtamoxifen‐independent recombination

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Last Updated: May 13, 2026

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Published on: January 8, 2015

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • The CreER-loxP system is crucial for genetic cell lineage tracing and conditional gene studies.
  • Unintended labeling of non-target cells by conventional CreER-loxP systems complicates data interpretation.

Purpose of the Study:

  • To engineer an improved CreER-loxP system with enhanced specificity and reduced background activity.
  • To minimize spontaneous nuclear translocation and nonspecific labeling in the absence of tamoxifen induction.

Main Methods:

  • Fusion of a membrane-localization motif to the C-terminus of CreER.
  • Development of a membrane-tethered CreER-loxP (mCreER-loxP) system.
  • Assessment of tamoxifen-induced nuclear translocation and recombination efficiency.

Main Results:

  • The mCreER-loxP system significantly reduced nonspecific labeling without tamoxifen.
  • Comparable recombination efficiency to conventional CreER-loxP was maintained upon induction.
  • Enhanced performance was observed under high expression and extended duration conditions in vitro.
  • Reduced cytotoxicity of mCreER compared to standard CreER was noted.

Conclusions:

  • The mCreER-loxP system offers improved temporal and spatial precision for genetic cell lineage tracing.
  • This engineered system provides a more reliable tool for genetic engineering applications.
  • The membrane-tethering strategy enhances the specificity and safety of CreER-loxP systems.