|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.)|
|Pagination||71 - 88|
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.
|Short Title||Methods in Molecular Biology (Clifton, N.J.)|