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A transcription activator-like effector toolbox for genome engineering.

Publication TypeJournal Article
AuthorsSanjana, NE, Cong L., Zhou Y., Cunniff MM, Feng G., and Zhang F.
AbstractTranscription activator-like effectors (TALEs) are a class of naturally occurring DNA-binding proteins found in the plant pathogen Xanthomonas sp. The DNA-binding domain of each TALE consists of tandem 34-amino acid repeat modules that can be rearranged according to a simple cipher to target new DNA sequences. Customized TALEs can be used for a wide variety of genome engineering applications, including transcriptional modulation and genome editing. Here we describe a toolbox for rapid construction of custom TALE transcription factors (TALE-TFs) and nucleases (TALENs) using a hierarchical ligation procedure. This toolbox facilitates affordable and rapid construction of custom TALE-TFs and TALENs within 1 week and can be easily scaled up to construct TALEs for multiple targets in parallel. We also provide details for testing the activity in mammalian cells of custom TALE-TFs and TALENs using quantitative reverse-transcription PCR and Surveyor nuclease, respectively. The TALE toolbox described here will enable a broad range of biological applications.
Year of Publication2012
JournalNature protocols
Volume7
Issue1
Pages171-92
Date Published (YYYY/MM/DD)2012/01/05
ISSN Number1754-2189
DOI10.1038/nprot.2011.431
PubMedhttp://www.ncbi.nlm.nih.gov/pubmed/22222791?dopt=Abstract