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Updated: May 18, 2026

Identification of Functional Protein Regions Through Chimeric Protein Construction

Identification of Functional Protein Regions Through Chimeric Protein Construction

Published on: January 8, 2019

Chimeric TALE recombinases with programmable DNA sequence specificity.

Andrew C Mercer¹, Thomas Gaj, Roberta P Fuller

¹The Skaggs Institute for Chemical Biology and Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

Nucleic Acids Research

|September 29, 2012

Summary

This summary is machine-generated.

Researchers developed TALE recombinases (TALERs) for genome engineering. These TALERs offer a more versatile and accessible alternative to zinc-finger recombinases for precise DNA modification in various cell types.

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Last Updated: May 18, 2026

Identification of Functional Protein Regions Through Chimeric Protein Construction

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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

Area of Science:

Molecular Biology
Genome Engineering
Synthetic Biology

Background:

Site-specific recombinases are crucial for genome engineering.
Hyperactivated serine recombinases can be re-targeted using DNA-binding domains (DBDs).
Zinc-finger proteins (ZFPs) as DBDs face limitations in modularity and DNA binding.

Purpose of the Study:

To develop an alternative to ZFPs for engineering recombinases.
To create chimeric TALE recombinases (TALERs) using transcription activator-like effector (TALE) proteins.
To assess TALER efficiency and specificity in bacterial and mammalian cells.

Main Methods:

Engineered fusions between a hyperactivated catalytic domain (Gin invertase) and optimized TALE architecture.
Utilized a library of incrementally truncated TALE variants.
Tested TALER DNA modification in bacterial and mammalian cells.

Main Results:

Identified TALER fusions with efficiency and specificity comparable to zinc-finger recombinases.
Demonstrated TALERs' ability to recombine DNA in bacterial cells.
Confirmed TALERs function in mammalian cells.

Conclusions:

TALERs provide a versatile platform for engineering recombinases with expanded targeting capabilities.
The TALER architecture offers a promising alternative to ZFPs for genome engineering.
TALERs have significant potential applications in biotechnology and medicine.