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

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2015 Oct 8

PMID 26443215

Citations 6

Authors

Pratiksha I Thakore

Charles A Gersbach

Affiliations

Soon will be listed here.

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.

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