A TALE nuclease architecture for efficient genome editing
A TALE nuclease architecture for efficient genome editing
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DOI：
10.1038/nbt.1755
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发表时间：
2011-02-01
影响因子：
46.9
通讯作者：
Rebar, Edward J.
中科院分区：
文献类型：
--
作者：
Miller, Jeffrey C.;Tan, Siyuan;Rebar, Edward J.
在活细胞中裂解独特基因组序列的核酸酶可用于靶向基因编辑和诱变。在这里，我们制定了一种基于Xanthomonas的转录激活效应子（Tale）蛋白质生成此类试剂的策略。我们确定了与FOKI催化结构域相连的Tale截断变体，可有效切割DNA，并使用这些核酸酶在内源性人NTF3和CCR5基因内以高达25％的效率产生离散的编辑或小缺失。我们进一步表明，设计的故事可以调节内源性哺乳动物基因。这些研究证明了设计的故事转录因子和核酸酶在靶向调节和内源基因的修饰中有效地应用。
Nucleases that cleave unique genomic sequences in living cells can be used for targeted gene editing and mutagenesis. Here we develop a strategy for generating such reagents based on transcription activator-like effector (TALE) proteins from Xanthomonas. We identify TALE truncation variants that efficiently cleave DNA when linked to the catalytic domain of FokI and use these nucleases to generate discrete edits or small deletions within endogenous human NTF3 and CCR5 genes at efficiencies of up to 25%. We further show that designed TALEs can regulate endogenous mammalian genes. These studies demonstrate the effective application of designed TALE transcription factors and nucleases for the targeted regulation and modification of endogenous genes.