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Conservative Site-specific Recombination and Phase Variation

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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.
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Related Experiment Video

Updated: Apr 9, 2026

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

Efficient conditional knockout targeting vector construction using co-selection BAC recombineering (CoSBR).

Robert J Newman¹, Merone Roose-Girma¹, Søren Warming²

¹Genentech, Inc., Department of Molecular Biology, 1 DNA Way, South San Francisco, CA 94080, USA.

Nucleic Acids Research

|June 20, 2015

Summary

This summary is machine-generated.

A new co-selection strategy simplifies Bacterial Artificial Chromosome (BAC) modification. This method efficiently alters two BAC positions simultaneously, streamlining gene targeting vector creation.

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Last Updated: Apr 9, 2026

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

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Published on: January 8, 2015

Recombineering Homologous Recombination Constructs in Drosophila

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Published on: July 13, 2013

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Published on: March 12, 2018

Area of Science:

Molecular Biology
Genetics Engineering

Background:

Bacterial Artificial Chromosomes (BACs) are essential tools for cloning large DNA fragments.
Modifying BACs for gene targeting often involves multiple complex steps.

Purpose of the Study:

To develop a simplified and efficient method for BAC modification.
To enable simultaneous alteration of two distinct sites on a BAC.

Main Methods:

Co-selection BAC recombineering strategy.
Co-transformation with single-stranded DNA oligos and double-stranded selection cassettes.
Two-step process: BAC modification followed by retrieval.

Main Results:

Efficient simultaneous modification of two BAC positions.
Reduced DNA manipulation for creating gene targeting vectors.
Streamlined workflow for conditional knockout vector construction.

Conclusions:

Co-selection BAC recombineering offers a significant improvement in efficiency and simplicity.
This strategy facilitates faster generation of complex genetic constructs.