Microfluidic MeDIP-seq for low-input methylomic analysis of mammary tumorigenesis in mice.
Microfluidic MeDIP-seq for low-input methylomic analysis of mammary tumorigenesis in mice.
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微流控 MeDIP-seq 用于小鼠乳腺肿瘤发生的低输入甲基组学分析。
DOI：
10.1039/c8an02271b
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Lu,Chang
中科院分区：
文献类型：
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作者：
Zhu,Yan;Cao,Zhenning;Lu,Chang
使用小鼠研究肿瘤发生过程中的动态表观基因组变化通常需要使用少量组织样本分析表观基因组。常规的表观基因组测试不支持这种分析，因为这些测定需要大量的材料。在这项研究中，我们开发了一种超灵敏的基于微流体的甲基化DNA免疫沉淀，然后是下一代测序（MeDIP-seq）技术，用于使用低至0.5 ng DNA（或100个细胞）的1.5 h芯片上免疫沉淀过程来分析甲基化组。这项技术使我们能够在乳腺癌发展的不同阶段检查C3（1）/SV 40 T抗原转基因小鼠模型中的全基因组DNA甲基化。利用我们的数据，我们确定了不同时期癌症发展的差异甲基化区域及其相关基因。我们的研究结果表明，独特的甲基化组学特征在不同的肿瘤发展阶段。
Studies of dynamic epigenomic changes during tumorigenesis using mice often require profiling epigenomes using a tiny quantity of tissue samples. Conventional epigenomic tests do not support such analysis due to the large amount of materials required by these assays. In this study, we developed an ultrasensitive microfluidics-based methylated DNA immunoprecipitation followed by next-generation sequencing (MeDIP-seq) technology for profiling methylomes using as little as 0.5 ng DNA (or ∼100 cells) with 1.5 h on-chip process for immunoprecipitation. This technology enabled us to examine genome-wide DNA methylation in a C3(1)/SV40 T-antigen transgenic mouse model during different stages of mammary cancer development. Using our data, we identified differentially methylated regions and their associated genes in different periods of cancer development. Our results showed that unique methylomic features were presented in various tumor developmental stages.