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Related Experiment Video

Updated: Feb 6, 2026

Molecular Evolution of the Tre Recombinase

Molecular Evolution of the Tre Recombinase

Published on: May 29, 2008

Directed Evolution of Targeted Recombinases for Genome Engineering.

Shannon J Sirk^1,2,3

¹Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA. sirk@illinois.edu.

Methods in Molecular Biology (Clifton, N.J.)

|August 30, 2018

Summary

This study details creating hybrid recombinase enzymes for precise genome engineering. These engineered enzymes allow for targeted DNA modifications at specific genomic locations.

Keywords:

Directed evolution Genetic engineering Genome editing Protein engineering Rational design Recombinase Site-specific genomic modification Targeted integration

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Published on: February 26, 2019

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Last Updated: Feb 6, 2026

Molecular Evolution of the Tre Recombinase

Molecular Evolution of the Tre Recombinase

Published on: May 29, 2008

Principles of Site-Specific Recombinase SSR Technology

Principles of Site-Specific Recombinase SSR Technology

Published on: May 29, 2008

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Published on: February 26, 2019

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Genome engineering is crucial for basic and clinical research.
Site-specific recombinases facilitate genome modification via DNA rearrangements.
Small serine recombinases offer modularity for custom genome targeting.

Purpose of the Study:

To describe the construction, selection, and validation of hybrid recombinase catalytic domains.
To enable targeted genome engineering at investigator-defined loci.
To leverage modular recombinase structure for novel genome modification tools.

Main Methods:

Designing hybrid recombinases by replacing native DNA-binding domains with targeting modules (e.g., zinc fingers, TALEs, inactivated Cas9).
Reprogramming the catalytic domain of serine recombinases for custom DNA sequence recognition.
Developing protocols for constructing, selecting, and validating these engineered enzymes.

Main Results:

Successful creation of hybrid recombinase catalytic domains.
Demonstration of reprogrammed catalytic domains for specific genomic targeting.
Validation of engineered recombinases for targeted genome engineering applications.

Conclusions:

Hybrid recombinases represent a powerful tool for precise genome engineering.
The modular nature of serine recombinases allows for versatile targeting strategies.
This work facilitates advanced applications in basic and clinical research through custom genome modification.