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| Status |
Public on Jan 24, 2018 |
| Title |
Single-cell nanobiopsy reveals compartmentalization of mRNAs within neuronal cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
In highly polarized cells such as neurons, compartmentalization of mRNA and of local protein synthesis enables remarkably fast, precise, and local responses to external stimuli. These responses are highly important for neuron growth cone guidance, synapse formation, and regeneration following injury. Because an altered spatial distribution of mRNA can result in mental retardation or neurodegenerative diseases, subcellular transcriptome analysis of neurons could be a useful tool for studying these conditions, but current techniques, such as in situ hybridization, bulk microarray, or RNA-Seq, impose tradeoffs between spatial resolution and multiplexing. To obtain a comprehensive analysis of the cell body versus neurite transcriptome from the same neuron, we have recently developed a label-free, single-cell nanobiopsy platform based on scanning ion conductance microscopy (SICM), that uses electrowetting within a quartz nanopipette to extract cellular material from living cells with minimal disruption of the cellular membrane and milieu. In this study, we used this platform to collect samples from the cell bodies and neurites of human neurons and analyzed the mRNA pool with multiplex RNA-Seq. The minute volume of a nanobiopsy sample allowed us to extract samples from several locations in the same cell and to map the various mRNA species to specific subcellular locations. In addition to previously identified transcripts, we discovered new sets of mRNAs localizing to neurites, including nuclear genes such as Eomes and Nap1l3. In summary, our single-neuron nanobiopsy analysis provides opportunities to improve our understanding of intracellular mRNA transport and local protein composition in neuronal growth, connectivity, and function.
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| Overall design |
Analysis of mRNA compartmentalization in cell bodies and neurites of human iPS-derived neuronal cells by nanobiopsy sampling and RNA-Sequencing.
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| Contributor(s) |
Tóth EN, Lohith A, Fukamizu A, Pourmand N |
| Citation(s) |
29378846 |
| Submission date |
Sep 05, 2017 |
| Last update date |
Jul 25, 2021 |
| Contact name |
Nader Pourmand |
| E-mail(s) |
pourmand@soe.ucsc.edu
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| Organization name |
University of California at Santa Cruz
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| Department |
Department of Biomolecular Engineering
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| Lab |
Pourmand Laboratory
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| Street address |
1156 High Street
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| City |
Santa Cruz |
| State/province |
CA |
| ZIP/Postal code |
95064 |
| Country |
USA |
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| Platforms (1) |
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| Samples (44)
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| Relations |
| BioProject |
PRJNA401740 |
| SRA |
SRP117061 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE103461_RAW.tar |
380.0 Kb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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