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Nat Protoc DOI:10.1038/nprot.2011.431

A transcription activator-like effector toolbox for genome engineering.

Publication TypeJournal Article
Year of Publication2012
AuthorsSanjana, NE, Cong, L, Zhou, Y, Cunniff, MM, Feng, G, Zhang, F
JournalNat Protoc
Date Published2012 Jan 05
KeywordsAnimals, Gene Expression Regulation, Genetic Engineering, Genomics, HEK293 Cells, Humans, Tandem Repeat Sequences, Transcription Factors, Xanthomonas

Transcription 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.


Alternate JournalNat Protoc
PubMed ID22222791
PubMed Central IDPMC3684555
Grant ListR01 NS073124 / NS / NINDS NIH HHS / United States
/ / Howard Hughes Medical Institute / United States