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

Biological Procedures Online

Claudia Uhde-Stone²  Joseph Huang¹  Biao Lu¹ 

[1] System Biosciences (SBI), 265 North Whisman Road, Mountain View CA 94043, USA

[2] Departments of Biological Sciences, California State University, East Bay, Hayward CA 94542, USA

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

Others  :  793191
DOI  :  10.1186/1480-9222-14-8

 received in 2012-07-20, accepted in 2012-07-22,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Uhde-Stone et al.; licensee BioMed Central Ltd.

【 图 表 】

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