Journal
ACS CHEMICAL NEUROSCIENCE
Volume 9, Issue 7, Pages 1858-1865Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acschemneuro.8b00197
Keywords
Nascent RNA; transcriptome-wide profiling; neuron; CuAAC; UPRT
Funding
- UQ Graduate School Scholarship
- UQ Development Award
- NSERC Postgraduate Award
- Westpac Future Leaders Scholarship
- ARC Discovery Project grant [DP180102998]
- National Institute of Mental Health R21 grant [1R21MH113062]
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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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