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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...
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...
Exon Recombination02:32

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon has three reading...
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
Gene Conversion02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...

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Recombineering Homologous Recombination Constructs in Drosophila
14:23

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

Recombination system based on cre alpha complementation and leucine zipper fusions.

Azadeh Seidi1, Masayasu Mie, Eiry Kobatake

  • 1Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.

Applied Biochemistry and Biotechnology
|November 13, 2008
PubMed
Summary

This study enhances a cre recombinase system for more efficient DNA recombination in mammalian cells. By fusing cre fragments to self-heterodimerizing proteins, the system

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CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion
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CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

Published on: June 25, 2017

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Last Updated: Jun 28, 2026

Recombineering Homologous Recombination Constructs in Drosophila
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Published on: July 13, 2013

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion
07:37

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

Published on: June 25, 2017

Area of Science:

  • Molecular Biology
  • Genetic Engineering

Background:

  • Previous cre recombinase systems relied on transient expression and protein transduction.
  • These systems aimed for controllable and less invasive recombination in mammalian cells.

Purpose of the Study:

  • To develop a more efficient cre recombinase complementation system.
  • To increase the association efficiency of cre recombinase fragments (alpha and beta).

Main Methods:

  • Fused cre fragments (alpha and beta) to leucine zipper motifs (bJun and bFos) from the AP-1 transcription factor.
  • Utilized the self-heterodimerizing properties of bJun and bFos to enhance fragment reassociation.

Main Results:

  • Achieved increased reassociation efficiency of the cre fragments.
  • Developed a recombination system that is two times more efficient than the previous study.

Conclusions:

  • The fusion of cre fragments to self-heterodimerizing proteins significantly improves recombination efficiency.
  • This enhanced system offers a more effective tool for genetic engineering in mammalian cells.