期刊
NATURE BIOTECHNOLOGY
卷 29, 期 2, 页码 149-U90出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt.1775
关键词
-
资金
- Harvard Society of Fellows
- National Human Genome Research Institute Center for Excellence in Genomics Science [P50 HG003170]
- Department of Energy Genomes to Life [DE-FG02-02ER63445]
- Defense Advanced Research Projects Agency [W911NF-08-1-0254]
- Wyss Institute for Biologically Inspired Engineering
- National Institutes of Health [R01 NS073124-01]
- European School of Molecular Medicine
- Church and Arlotta laboratories
- U.S. Department of Energy (DOE) [DE-FG02-02ER63445] Funding Source: U.S. Department of Energy (DOE)
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.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据