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Related Experiment Video

Updated: May 15, 2026

Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters

Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters

Published on: November 26, 2013

A do-it-yourself protocol for simple transcription activator-like effector assembly.

Claudia Uhde-Stone¹, Nilang Gor, Tiffany Chin

¹System Biosciences (SBI), 265 North Whisman Rd, Mountain View, CA, 94043, USA. BLu@systembio.com.

Biological Procedures Online

|January 16, 2013

Summary

This summary is machine-generated.

This study presents a simplified, do-it-yourself method for assembling transcription activator-like effectors (TALEs) and TALE nucleases (TALENs) in just two days. This accessible TALE assembly system broadens the use of TALE technology for genetic engineering.

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Area of Science:

Molecular Biology
Genome Engineering
Synthetic Biology

Background:

Transcription activator-like effectors (TALEs) are versatile tools for genetic and epigenetic manipulation due to their predictable binding and specificity.
Current TALE assembly methods are often expensive, time-consuming, and require specialized expertise, limiting their widespread adoption.

Purpose of the Study:

To develop a streamlined and accessible method for custom TALE assembly.
To enable individual researchers to efficiently generate TALE-based tools for genome engineering.

Main Methods:

A simplified Golden Gate assembly protocol was developed using pre-made, quality-controlled TALE monomers.
The protocol was optimized for a two-day assembly process involving four thermocycling reactions.
Vector sets for generating both TALE-transcription factors (TALE-TFs) and TALE nucleases (TALENs) were provided.

Main Results:

A rapid, two-day TALE assembly method was established, eliminating complex setup procedures.
The system successfully generated various TALENs and TALE-TFs, confirmed by DNA sequencing.
An assembled TALE-TF demonstrated transactivation of a luciferase reporter, and a TALEN pair effectively cleaved its target DNA.

Conclusions:

A validated do-it-yourself system for assembling TALENs and TALE-TFs within two days has been developed.
This simplified TALE assembly approach, coupled with versatile vector options, is expected to significantly increase the accessibility and application of TALE technology.