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Conservative Site-specific Recombination and Phase Variation

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...

Gene Conversion

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...

Homologous Recombination

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...

Crossing Over

Crossing Over

Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to allelic diversity in the newly formed daughter cells. In humans, crossing over produces genetically distinct haploid egg and sperm cells that undergo fertilization to produce unique offspring. Before cell division starts, the germ cell’s chromosome(s) undergo duplication in the S phase of the cell cycle. As the cells enter prophase I,...

Viral Recombination

Viral Recombination

Cells are sometimes infected by more than one virus at once. When two viruses disassemble to expose their genomes for replication in the same cell, similar regions of their genomes can pair together and exchange sequences in a process called recombination. Alternatively, viruses with segmented genomes can swap segments in a process called reassortment.

Exon Recombination

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...

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Related Experiment Video

Updated: May 30, 2025

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Genome recombination on demand.

Marta Seczynska¹, Lars M Steinmetz^1,2,3

¹Department of Genetics, School of Medicine, Stanford University, Stanford, CA, USA.

Science (New York, N.Y.)

|January 30, 2025

Summary

This summary is machine-generated.

Large genome rearrangements can now be created efficiently in mammalian cells. This breakthrough enables large-scale genomic studies and applications in various biological fields.

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Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

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Last Updated: May 30, 2025

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy

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Published on: June 25, 2013

Recombineering Homologous Recombination Constructs in Drosophila

Recombineering Homologous Recombination Constructs in Drosophila

Published on: July 13, 2013

Area of Science:

Genomics
Cell Biology
Molecular Biology

Background:

Genome rearrangements are crucial for cellular function and evolution.
Previous methods for generating large genome rearrangements were limited in scale and efficiency.

Purpose of the Study:

To develop a scalable method for generating large genome rearrangements in mammalian cells.
To enable high-throughput studies of genome instability and DNA repair.

Main Methods:

Utilized CRISPR-Cas9 gene editing technology.
Designed specific guide RNAs to induce double-strand breaks at desired genomic locations.
Employed DNA repair pathways to facilitate rearrangements.

Main Results:

Successfully generated a variety of large genome rearrangements, including deletions, inversions, and translocations.
Demonstrated scalability of the method across different mammalian cell types.
Validated the accuracy and efficiency of the generated rearrangements.

Conclusions:

The developed method provides a powerful tool for large-scale genome engineering in mammalian cells.
This advancement will accelerate research in areas such as cancer genomics, developmental biology, and synthetic biology.