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ACS Chem Neurosci. 2018 Jul 18;9(7):1858-1865. doi: 10.1021/acschemneuro.8b00197. Epub 2018 Jun 15.

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

Author information

1
Cognitive Neuroepigenetics Laboratory , Queensland Brain Institute, The University of Queensland , Brisbane , Queensland 4072 , Australia.

Abstract

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.

KEYWORDS:

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

PMID:
29874042
PMCID:
PMC6272126
[Available on 2019-07-18]
DOI:
10.1021/acschemneuro.8b00197

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