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

ABSTRACT: 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.

SUBMITTER: Thakore PI

PROVIDER: S-EPMC4922632 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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

Thakore Pratiksha I PI Gersbach Charles A CA

Methods in molecular biology (Clifton, N.J.) 20160101

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 ...[more]

PMID: 26443215

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