Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors.
Publication
, Journal Article
Thakore, PI; Gersbach, CA
Published in: Methods in molecular biology (Clifton, N.J.)
January 2016
Transcription activator-like effectors (TALEs) are modular DNA-binding proteins that can be fused to a variety of effector domains to regulate the epigenome. Nucleotide recognition by TALE monomers follows a simple cipher, making this a powerful and versatile method to activate or repress gene expression. Described here are methods to design, assemble, and test TALE transcription factors (TALE-TFs) for control of endogenous gene expression. In this protocol, TALE arrays are constructed by Golden Gate cloning and tested for activity by transfection and quantitative RT-PCR. These methods for engineering TALE-TFs are useful for studies in reverse genetics and genomics, synthetic biology, and gene therapy.
Duke Scholars
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Published In
Methods in molecular biology (Clifton, N.J.)
DOI
EISSN
1940-6029
ISSN
1064-3745
Publication Date
January 2016
Volume
1338
Start / End Page
71 / 88
Related Subject Headings
- Trans-Activators
- Synthetic Biology
- Promoter Regions, Genetic
- Humans
- HEK293 Cells
- Genomics
- Genetic Vectors
- Genetic Engineering
- Gene Targeting
- Developmental Biology
Citation
APA
Chicago
ICMJE
MLA
NLM
Thakore, P. I., & Gersbach, C. A. (2016). Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors. Methods in Molecular Biology (Clifton, N.J.), 1338, 71–88. https://doi.org/10.1007/978-1-4939-2932-0_7
Thakore, Pratiksha I., and Charles A. Gersbach. “Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors.” Methods in Molecular Biology (Clifton, N.J.) 1338 (January 2016): 71–88. https://doi.org/10.1007/978-1-4939-2932-0_7.
Thakore PI, Gersbach CA. Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors. Methods in molecular biology (Clifton, NJ). 2016 Jan;1338:71–88.
Thakore, Pratiksha I., and Charles A. Gersbach. “Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors.” Methods in Molecular Biology (Clifton, N.J.), vol. 1338, Jan. 2016, pp. 71–88. Epmc, doi:10.1007/978-1-4939-2932-0_7.
Thakore PI, Gersbach CA. Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors. Methods in molecular biology (Clifton, NJ). 2016 Jan;1338:71–88.
Published In
Methods in molecular biology (Clifton, N.J.)
DOI
EISSN
1940-6029
ISSN
1064-3745
Publication Date
January 2016
Volume
1338
Start / End Page
71 / 88
Related Subject Headings
- Trans-Activators
- Synthetic Biology
- Promoter Regions, Genetic
- Humans
- HEK293 Cells
- Genomics
- Genetic Vectors
- Genetic Engineering
- Gene Targeting
- Developmental Biology