Bioorthogonal Metabolic Labeling of Nascent RNA in Neurons Improves the Sensitivity of Transcriptome-Wide Profiling,ACS Chemical Neuroscience

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Bioorthogonal Metabolic Labeling of Nascent RNA in Neurons Improves the Sensitivity of Transcriptome-Wide Profiling
ACS Chemical Neuroscience ( IF 4.1 ) Pub Date : 2018-06-06 00:00:00 , DOI: 10.1021/acschemneuro.8b00197
Esmi L Zajaczkowski ₁ , Qiong-Yi Zhao ₁ , Zong Hong Zhang ₁ , Xiang Li ₁ , Wei Wei ₁ , Paul R Marshall ₁ , Laura J Leighton ₁ , Sarah Nainar , Chao Feng , Robert C Spitale , Timothy W Bredy ₁

Affiliation

Transcriptome-wide expression profiling of neurons has provided important insights into the underlying molecular mechanisms and gene expression patterns that transpire during learning and memory formation. However, there is a paucity of tools for profiling stimulus-induced RNA within specific neuronal cell populations. A bioorthogonal method to chemically label nascent (i.e., newly transcribed) RNA in a cell-type-specific and temporally controlled manner, which is also amenable to bioconjugation via click chemistry, was recently developed and optimized within conventional immortalized cell lines. However, its value within a more fragile and complicated cellular system such as neurons, as well as for transcriptome-wide expression profiling, has yet to be demonstrated. Here, we report the visualization and sequencing of activity-dependent nascent RNA derived from neurons using this labeling method. This work has important implications for improving transcriptome-wide expression profiling and visualization of nascent RNA in neurons, which has the potential to provide valuable insights into the mechanisms underlying neural plasticity, learning, and memory.

中文翻译：

神经元新生 RNA 的生物正交代谢标记提高了全转录组分析的灵敏度

神经元的全转录组表达谱为学习和记忆形成过程中发生的潜在分子机制和基因表达模式提供了重要的见解。然而，用于分析特定神经元细胞群内刺激诱导的 RNA 的工具很少。最近开发了一种生物正交方法，以细胞类型特异性和时间控制的方式化学标记新生（即新转录的）RNA，该方法也适合通过点击化学进行生物缀合，并且在传统的永生化细胞系中进行了优化。然而，它在神经元等更脆弱和复杂的细胞系统中以及在转录组范围内的表达谱分析中的价值尚未得到证实。在这里，我们报告了使用这种标记方法对来自神经元的活性依赖性新生 RNA 进行可视化和测序。这项工作对于改善神经元中新生 RNA 的转录组表达谱和可视化具有重要意义，这有可能为神经可塑性、学习和记忆的机制提供有价值的见解。

更新日期：2018-06-06

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