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- Title
Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription.
- Authors
Feng Zhang; Le Cong; Lodato, Simona; Kosuri, Sriram; Church, George M.; Arlotta, Paola
- Abstract
The ability to direct functional proteins to specific DNA sequences is a long-sought goal in the study and engineering of biological processes. Transcription activator-like effectors (TALEs) from Xanthomonas sp. are site-specific DNA-binding proteins that can be readily designed to target new sequences. Because TALEs contain a large number of repeat domains, it can be difficult to synthesize new variants. Here we describe a method that overcomes this problem. We leverage codon degeneracy and type IIs restriction enzymes to generate orthogonal ligation linkers between individual repeat monomers, thus allowing full-length, customized, repeat domains to be constructed by hierarchical ligation. We synthesized 17 TALEs that are customized to recognize specific DNA-binding sites, and demonstrate that they can specifically modulate transcription of endogenous genes (SOX2 and KLF4) in human cells.
- Publication
Nature Biotechnology, 2011, Vol 29, Issue 2, p149
- ISSN
1087-0156
- Publication type
Academic Journal
- DOI
10.1038/nbt.1775