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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.

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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

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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