In vivo site-specific DNA methylation with a designed sequence-enabled DNA methylase
In vivo site-specific DNA methylation with a designed sequence-enabled DNA methylase
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DOI：
10.1021/ja0705588
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发表时间：
2007-07-18
影响因子：
15
通讯作者：
Barbas, Carlos F., III
中科院分区：
文献类型：
--
作者：
Nomura, Wataru;Barbas, Carlos F., III
DNA的共价修饰，如胞嘧啶甲基化，可以诱导可遗传的基因沉默。如果表观遗传修饰可以特异性靶向，那么转录治疗的新方法应该会产生。为了解决这一挑战，我们试图设计甲基转移酶，通过调整序列使能的组装策略，仅在所需的位点起作用。我们的分裂DNA甲基化酶在活细胞中进行位点特异性CpG甲基化，当在靶位点适当组装时，没有任何背景甲基化。这是序列使能酶重组方法在体内的首次成功应用。这种分裂酶重组策略将允许产生可编程的锌指甲基化酶，其作用于哺乳动物基因组中的任何特定CpG位点。可编程甲基化酶应该协调可遗传的基因沉默，并应在DNA标记方法和纳米技术中找到应用。
Covalent modification of DNA, such as cytosine methylation, can induce heritable gene silencing. If epigenetic modifications can be specifically targeted, new approaches to transcriptional therapy should result. To address this challenge we sought to design methyltransferases that would act only at a desired site by adapting the sequence-enabled assembly strategy. Our split DNA methylase performed site-specific CpG methylation in living cells without any background methylation when appropriately assembled at the target site. This is the first successful application of the sequence-enable enzyme reassembly approach in vivo. This split-enzyme reassembly strategy will allow creation of programmable zinc finger methylases that act at any specific CpG site in the mammalian genome. Programmable methylases should orchestrate heritable gene silencing and should find application in DNA tagging approaches and in nanotechnology.