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A robust dual reporter system to visualize and quantify gene expression mediated by transcription activator-like effectors

Claudia Uhde-Stone1, Joseph Huang2 and Biao Lu2*

Author Affiliations

1 Departments of Biological Sciences, California State University, East Bay, Hayward CA 94542, USA

2 System Biosciences (SBI), 265 North Whisman Road, Mountain View CA 94043, USA

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Biological Procedures Online 2012, 14:8 doi:10.1186/1480-9222-14-8

Published: 8 August 2012

Abstract

Background

Transcription activator-like effectors (TALEs) are a class of naturally occurring transcription effectors that recognize specific DNA sequences and modulate gene expression. The modularity of TALEs DNA binding domain enables sequence-specific perturbation and offers broad applications in genetic and epigenetic studies. Although the efficient construction of TALEs has been established, robust functional tools to assess their functions remain lacking.

Results

We established a dual reporter system that was specifically designed for real-time monitoring and quantifying gene expression mediated by TALEs. We validated both sensitivity and specificity of this dual-reporter system in mammalian cells, and demonstrated that this dual reporter system is robust and potentially amenable to high throughput (HTP) applications.

Conclusion

We have designed, constructed and validated a novel dual reporter system for assessing TALE mediated gene regulations. This system offers a robust and easy-to- use tool for real-time monitoring and quantifying gene expression in mammalian cells.

Keywords:
Dual reporter; Gene editing; Transcription activator-like effector; Green fluorescent protein; Firefly luciferase