4.6 Article

Bioorthogonal Metabolic Labeling of Nascent RNA in Neurons Improves the Sensitivity of Transcriptome-Wide Profiling

期刊

ACS CHEMICAL NEUROSCIENCE
卷 9, 期 7, 页码 1858-1865

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acschemneuro.8b00197

关键词

Nascent RNA; transcriptome-wide profiling; neuron; CuAAC; UPRT

资金

  1. UQ Graduate School Scholarship
  2. UQ Development Award
  3. NSERC Postgraduate Award
  4. Westpac Future Leaders Scholarship
  5. ARC Discovery Project grant [DP180102998]
  6. National Institute of Mental Health R21 grant [1R21MH113062]

向作者/读者索取更多资源

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.

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