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Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

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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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Updated: Mar 6, 2026

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion
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A new location to split Cre recombinase for protein fragment complementation.

Maryam Rajaee1,2, David W Ow1

  • 1Plant Gene Engineering Center, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, China.

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|March 21, 2017
PubMed
Summary

A split Cre recombinase system was developed to overcome issues with maintaining high Cre-expressing plant lines. Splitting Cre between Lys244 and Asn245 reconstitutes activity comparable to wild-type Cre.

Keywords:
alpha-complementationgene stackingprotein fragment complementationsite-specific recombination

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Area of Science:

  • Molecular Biology
  • Plant Biotechnology
  • Genetic Engineering

Background:

  • Recombinase-mediated gene stacking relies on Cre recombinase for DNA removal.
  • Introducing Cre via hybridization can lead to issues maintaining stable, high-expressing plant lines.
  • High Cre activity may negatively impact normal chromosome functions.

Purpose of the Study:

  • To develop a split Cre recombinase system for improved gene stacking.
  • To identify new locations for splitting Cre into complementing fragments.
  • To maintain the functional freedom of recombinase-mediated gene stacking systems.

Main Methods:

  • Testing four novel locations for splitting the Cre recombinase.
  • Utilizing protein fragment complementation assays.
  • Assessing reconstituted Cre activity against wild-type Cre.

Main Results:

  • Four new potential split sites for Cre recombinase were investigated.
  • Splitting Cre between Lys244 and Asn245 yielded functional complementation.
  • The reconstituted Cre activity was comparable to wild-type Cre activity.

Conclusions:

  • A split Cre system offers a viable alternative to constitutive Cre expression in plants.
  • The Lys244-Asn245 split site effectively reconstitutes Cre recombinase function.
  • This approach enhances the utility of recombinase-mediated gene stacking.