CRISPR-based targeting of DNA methylation in Arabidopsis thaliana by a bacterial CG-specific DNA methyltransferase.
CRISPR-based targeting of DNA methylation in Arabidopsis thaliana by a bacterial CG-specific DNA methyltransferase.
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以CRISPR为基础的细菌CG特异性DNA甲基转移酶对拟南芥DNA甲基化的靶向性研究
DOI：
10.1073/pnas.2125016118
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发表时间：
2021-06-08
影响因子：
11.1
通讯作者：
Jacobsen SE
中科院分区：
文献类型：
--
作者：
Ghoshal B;Picard CL;Vong B;Feng S;Jacobsen SE
位点特异性修饰表观遗传标记，如DNA甲基化，以调节基因表达是一种独特的方法，以提高重要的经济作物性状。这种方法允许在没有初始转基因诱导剂的情况下在后代中维持引入的修饰，并且主要依赖于cg特异性序列背景下胞嘧啶的甲基化。我们开发了一种基于细菌甲基转移酶和CRISPR-Cas9平台的靶向DNA甲基化工具，直接甲基化拟南芥CG位点的胞嘧啶。这些工具扩展了目前可用的基于crispr的靶向DNA甲基化工具，并为在植物中建立可遗传的靶向DNA甲基化提供了一种方法。基于crispr的表观遗传标记靶向修饰（如DNA胞嘧啶甲基化）是调控基因表达及其相关表型的重要策略。虽然植物在所有序列背景下都有DNA甲基化（CG， CHG， CHH，其中H = A， T， C），但对称CG背景下的甲基化对基因沉默尤为重要，并通过有丝分裂和减数分裂细胞分裂非常有效地维持。因此，可以直接将CG甲基化添加到特定位点的工具是非常需要的，但目前在植物中缺乏。在这里，我们开发了两种基于crispr的植物cg特异性靶向DNA甲基化系统，使用细菌cg特异性DNA甲基化酶MQ1的变体，活性降低，但特异性高。我们证明了MQ1添加的甲基化是高度靶向特异性的，并且可以在没有效应物的情况下遗传地维持。这些工具在作物工程和植物遗传研究中都很有价值。
Site-specific modification of epigenetic marks such as DNA methylation to regulate gene expression is a unique approach to enhance economically important crop traits. This approach allows for the maintenance of the introduced modifications in the absence of the initial transgene inducer in subsequent generations and relies largely on methylation of cytosines in the CG-specific sequence context. We have developed a targeted DNA methylation tool based on a bacterial methyltransferase and the CRISPR-Cas9 platform to directly methylate cytosines at CG sites in Arabidopsis. These tools expand the currently available CRISPR-based targeted DNA methylation tools and provide an approach for the establishment of heritable targeted DNA methylation in plants. CRISPR-based targeted modification of epigenetic marks such as DNA cytosine methylation is an important strategy to regulate the expression of genes and their associated phenotypes. Although plants have DNA methylation in all sequence contexts (CG, CHG, CHH, where H = A, T, C), methylation in the symmetric CG context is particularly important for gene silencing and is very efficiently maintained through mitotic and meiotic cell divisions. Tools that can directly add CG methylation to specific loci are therefore highly desirable but are currently lacking in plants. Here we have developed two CRISPR-based CG-specific targeted DNA methylation systems for plants using a variant of the bacterial CG-specific DNA methyltransferase MQ1 with reduced activity but high specificity. We demonstrate that the methylation added by MQ1 is highly target specific and can be heritably maintained in the absence of the effector. These tools should be valuable both in crop engineering and in plant genetic research.
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DOI：
10.1038/nrm4043
发表时间：
2015-09
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Du J;Johnson LM;Jacobsen SE;Patel DJ
通讯作者：
Patel DJ
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
16.8
作者：
Stroud H;Do T;Du J;Zhong X;Feng S;Johnson L;Patel DJ;Jacobsen SE
通讯作者：
Jacobsen SE
DOI：
10.1073/pnas.1716945115
发表时间：
2018-02-27
影响因子：
11.1
作者：
Gallego-Bartolomé J;Gardiner J;Liu W;Papikian A;Ghoshal B;Kuo HY;Zhao JM;Segal DJ;Jacobsen SE
通讯作者：
Jacobsen SE
影响因子：
9.5
作者：
Thorvaldsdóttir H;Robinson JT;Mesirov JP
通讯作者：
Mesirov JP