Title | Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors. |
Publication Type | Journal Article |
Year of Publication | 2016 |
Authors | PI Thakore, and CA Gersbach |
Journal | Methods in molecular biology (Clifton, N.J.) |
Volume | 1338 |
Start Page | 71 |
Pagination | 71 - 88 |
Date Published | 01/2016 |
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. |
DOI | 10.1007/978-1-4939-2932-0_7 |
Short Title | Methods in molecular biology (Clifton, N.J.) |