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Engineering Customized TALENs Using the Platinum Gate TALEN Kit.

Tetsushi Sakuma1, Takashi Yamamoto2

  • 1Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8526, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|October 8, 2015
PubMed
Summary
This summary is machine-generated.

Golden Gate assembly is a popular method for creating transcription activator-like effector nucleases (TALENs). This study details protocols for TALEN construction using the Platinum Gate TALEN Kit, enabling the generation of highly active TALENs.

Keywords:
4-Module assemblyGolden Gate assemblyModular assemblyPlatinum Gate TALEN KitPlatinum TALEN

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Area of Science:

  • Molecular Biology
  • Gene Editing Technologies

Background:

  • Transcription activator-like effector nucleases (TALENs) are powerful tools for targeted genome modification.
  • Golden Gate assembly is a widely adopted and well-characterized method for TALEN construction.
  • Existing methods for TALEN assembly can be labor-intensive and require multiple purification steps.

Purpose of the Study:

  • To describe protocols for constructing TALENs using Golden Gate assembly.
  • To introduce the Platinum Gate TALEN Kit for efficient TALEN generation.
  • To enable the creation of highly active Platinum TALENs.

Main Methods:

  • Utilizing the Golden Gate assembly principle for DNA-binding repeat concatemerization.
  • Employing a single-tube, cycling reaction of digestion and ligation.
  • Implementing the Platinum Gate TALEN Kit for streamlined TALEN construction.

Main Results:

  • Demonstration of PCR-, fragmentation-, and purification-free DNA-binding repeat assembly.
  • Successful generation of TALEN constructs using the described protocols.
  • Generation of highly active Platinum TALENs.

Conclusions:

  • Golden Gate assembly provides an efficient strategy for TALEN construction.
  • The Platinum Gate TALEN Kit simplifies and enhances the TALEN generation process.
  • This method facilitates the production of active TALENs for various research applications.