Single-neuron transcriptome and methylome sequencing for epigenomic analysis of aging.
Single-neuron transcriptome and methylome sequencing for epigenomic analysis of aging.
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DOI：
10.1007/978-1-62703-556-9_21
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
Kohn, Andrea B
中科院分区：
文献类型：
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作者：
Moroz, Leonid L;Kohn, Andrea B
转录和信号的巨大异质性是减缓我们对正常衰老和年龄相关疾病机制的理解进展的特征。这对于衰老的神经生物学是至关重要的，因为神经元群体的巨大多样性在实验设计中提出了巨大的挑战。在这里，我们介绍了Aplysia作为单细胞水平上衰老的基因组分析的模型，并提供了在衰老过程中单独识别的神经元的整合转录组和甲基组图谱的方案。这些单细胞RNA-SEQ和DNA甲基化分析(甲基捕获/甲基浓缩)与所有主要的下一代测序平台兼容(我们使用了Roche/454和Solid/Life Technologies作为例证)，并可用于将表观遗传特征与转录输出相结合。所描述的测序文库构建方案提供了来自单个细胞转录图谱的定量和定向信息。我们的结果还证实，在多倍体海兔神经元中，不同拷贝的DNA在DNA甲基化方面的行为相似。
Enormous heterogeneity in transcription and signaling is the feature that slows down progress in our understanding of the mechanisms of normal aging and age-related diseases. This is critical for neurobiology of aging where the enormous diversity of neuronal populations presents a significant challenge in experimental design. Here, we introduce Aplysia as a model for genomic analysis of aging at the single-cell level and provide protocols for integrated transcriptome and methylome profiling of individually identified neurons during the aging process. These single-cell RNA-seq and DNA methylation assays (methyl-capture/methyl enrichment) are compatible with all major next generation sequencing platforms (we used Roche/454 and SOLiD/Life Technologies as illustrative examples) and can be used to integrate an epigenetic signature with transcriptional output. The described sequencing library construction protocol provides both quantitative and directional information from transcriptional profiling of individual cells. Our results also confirm that different copies of DNA in polyploid Aplysia neurons behave similarly with respect to their DNA methylation.