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

TitleDesign, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors.
Publication TypeJournal Article
Year of Publication2016
AuthorsThakore, PI, and Gersbach, CA
JournalMethods in molecular biology (Clifton, N.J.)
Volume1338
Start Page71
Pagination71 - 88
Date Published01/2016
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.

DOI10.1007/978-1-4939-2932-0_7
Short TitleMethods in molecular biology (Clifton, N.J.)