Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Thakore, PI; Gersbach, CA

Published Date

  • January 2016

Published In

Volume / Issue

  • 1338 /

Start / End Page

  • 71 - 88

PubMed ID

  • 26443215

Pubmed Central ID

  • PMC4922632

Electronic International Standard Serial Number (EISSN)

  • 1940-6029

International Standard Serial Number (ISSN)

  • 1064-3745

Digital Object Identifier (DOI)

  • 10.1007/978-1-4939-2932-0_7

Language

  • eng