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Related Experiment Video

Updated: May 9, 2026

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
08:31

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Published on: February 5, 2021

A simple and efficient method for assembling TALE protein based on plasmid library.

Zhiqiang Zhang1, Duo Li, Huarong Xu

  • 1College of Animal Science and Technology, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling, Shaan'xi, PR China.

Plos One
|July 11, 2013
PubMed
Summary
This summary is machine-generated.

We developed a simple, rapid, and cost-efficient method for assembling TALE repeat DNA sequences. This approach facilitates the creation of custom transcription activator-like effector nucleases (TALENs) and transcription factors (TALE-TFs) for genome engineering.

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Last Updated: May 9, 2026

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Area of Science:

  • Molecular Biology
  • Genomics
  • Synthetic Biology

Background:

  • Transcription activator-like effectors (TALEs) offer precise DNA targeting for genome engineering.
  • Assembling long TALE repeat arrays is a significant hurdle, limiting broader application.
  • Existing methods often require specialized equipment or expertise.

Purpose of the Study:

  • To present a straightforward and efficient method for constructing custom TALE nucleases (TALENs) and TALE transcription factors (TALE-TFs).
  • To overcome the challenges associated with assembling long TALE repeat arrays.

Main Methods:

  • Utilized a TALE repeat tetramer library (256 combinations) with unique primers for PCR amplification.
  • Assembled PCR products in a single digestion/ligation step.
  • Generated TALEN and TALE-TF constructs targeting specific mouse genes (Gt(ROSA)26Sor, Mstn, Oct4, c-Myc, Klf4, Sox2).

Main Results:

  • Successfully generated 12 TALE constructs (4 TALEN pairs, 4 TALE-TFs) within 3 days.
  • Achieved an average positive clone rate of 64% via colony PCR.
  • Demonstrated high success rates in sequencing, confirming construct integrity.

Conclusions:

  • The developed method is rapid, cost-efficient, and utilizes common laboratory resources.
  • This technique simplifies the generation of customized TALENs and TALE-TFs, promoting wider adoption of TALE technology.
  • The high success rate and ease of use make this a valuable tool for genome engineering applications.