Design, Assembly, and Characterization of TALE-Based Transcriptional Activators and Repressors.
Journal Article (Journal Article)
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.
Full Text
Duke Authors
Cited Authors
- Thakore, PI; Gersbach, CA
Published Date
- January 2016
Published In
Volume / Issue
- 1338 /
Start / End Page
- 71 - 88
PubMed ID
- 26443215
Pubmed Central ID
- PMC4922632
Electronic International Standard Serial Number (EISSN)
- 1940-6029
International Standard Serial Number (ISSN)
- 1064-3745
Digital Object Identifier (DOI)
- 10.1007/978-1-4939-2932-0_7
Language
- eng