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Related Experiment Video

Updated: May 4, 2026

Mouse Genome Engineering Using Designer Nucleases
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A single-chain TALEN architecture for genome engineering.

Ning Sun1, Huimin Zhao

  • 1Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

Molecular Biosystems
|December 17, 2013
PubMed
Summary

Researchers developed a single-chain TALEN (scTALEN) to simplify genome engineering. This novel TALEN architecture reduces protein payload and production costs for more efficient gene editing applications.

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Transcription-activator like effector nucleases (TALENs) are engineered DNA-binding proteins used for genome engineering.
  • TALENs function via dimerization of FokI nuclease domains at targeted DNA sites.
  • Delivering and expressing two large TALEN monomers can be challenging for some applications.

Purpose of the Study:

  • To develop a novel single-chain TALEN (scTALEN) architecture.
  • To overcome challenges associated with delivering and expressing multiple TALEN components.
  • To create a more efficient and cost-effective genome engineering tool.

Main Methods:

  • Designed a single-chain TALEN (scTALEN) by fusing two FokI nuclease domains with a polypeptide linker.
  • Utilized a 95 amino acid polypeptide linker isolated from a library via high-throughput screening.

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  • Tested the catalytic activity of scTALENs in yeast and human cells.

    • Main Results:

    • Demonstrated that scTALENs are catalytically active as monomers.
    • Confirmed functionality in both yeast and human cellular systems.
    • Showcased the potential for reduced protein payload and simplified design.

    Conclusions:

    • The novel scTALEN architecture offers a promising alternative to traditional TALEN systems.
    • scTALENs can simplify the design and reduce the production costs of genome engineering tools.
    • This approach facilitates broader implementation of TALEN technology in various applications.