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Related Experiment Video

Updated: Apr 20, 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

Genome engineering with custom recombinases.

Thomas Gaj¹, Carlos F Barbas¹

¹Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA; Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.

Methods in Enzymology

|November 16, 2014

Summary

This summary is machine-generated.

This study details creating custom hybrid recombinases for precise genome engineering. These tools enable targeted DNA modifications in mammalian cells, offering a modular approach for research and applications.

Keywords:

Genome engineering Protein engineering Recombinase TAL effector Zinc-finger

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Recombineering Homologous Recombination Constructs in Drosophila

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Published on: July 13, 2013

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Site-specific recombinases are essential for genetic research and engineering.
Hybrid recombinases combine serine recombinase catalytic domains with DNA-binding domains (zinc-finger or TAL effector).

Purpose of the Study:

To provide a protocol for generating novel hybrid recombinases with user-defined DNA targeting specificity.
To outline methods for site-specific integration of genetic material into targeted genomic loci using these engineered recombinases.

Main Methods:

Construction of hybrid recombinases by fusing catalytic and DNA-binding domains.
Generation of recombinases with tailored specificity for "plug-and-play" applications.
Implementation of site-specific integration strategies in mammalian cells.

Main Results:

Demonstration of a modular system for creating custom recombinases.
Successful generation of hybrid recombinases with specific targeting capabilities.
Establishment of methods for precise genomic modifications.

Conclusions:

The developed protocol facilitates the creation of versatile hybrid recombinases for diverse genome engineering tasks.
This approach enables precise, targeted genetic modifications in mammalian systems.
The modularity allows for rapid development of new recombinases with novel specificities.