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Status |
Public on Dec 07, 2017 |
Title |
Dissecting cell-type composition and activity-dependent transcriptional state in mammalian brains by massively parallel single-nucleus RNA-Seq |
Organism |
Mus musculus |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Massively parallel single-cell RNA sequencing can precisely resolve cellular diversity in a high-throughput manner at low cost, but unbiased isolation of intact single cells from complex tissues, such as adult mammalian brains, is challenging. Here, we integrate sucrose-gradient assisted nuclei purification with droplet microfluidics to develop a highly scalable single-nucleus RNA-Seq approach (sNucDrop-Seq), which is free of enzymatic dissociation and nuclei sorting. By profiling ~18,000 nuclei isolated from cortical tissues of adult mice, we demonstrate that sNucDrop-Seq not only accurately reveals neuronal and non-neuronal subtype composition with high sensitivity, but also enables in-depth analysis of transient transcriptional states driven by neuronal activity, at single-cell resolution, in vivo.
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Overall design |
Single-nucleus RNA sequencing analysis of adult mouse cerebral cortex
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Contributor(s) |
Hu P, Fabyanic E, Kwon D, Tang S, Zhou Z, Wu H |
Citation(s) |
29220646 |
NIH grant(s) |
Grant ID |
Grant title |
Affiliation |
Name |
DP2 HL142044 |
Charting oxygen-sensing gene regulatory network in cardiomyocytes through single-cell analysis and epigenome editing |
UNIVERSITY OF PENNSYLVANIA |
Hao Wu |
R00 HG007982 |
Genomic and functional analysis of active DNA demethylation in mammalian cells |
UNIVERSITY OF PENNSYLVANIA |
Hao Wu |
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Submission date |
Nov 08, 2017 |
Last update date |
May 15, 2019 |
Contact name |
Hao Wu |
E-mail(s) |
haowu2@pennmedicine.upenn.edu
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Phone |
215-573-9360
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Organization name |
University of Pennsylvania
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Department |
Genetics
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Lab |
Hao Wu
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Street address |
415 Curie Boulevard
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City |
Philadelphia |
State/province |
PA |
ZIP/Postal code |
19104 |
Country |
USA |
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Platforms (1) |
GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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Samples (19)
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Relations |
BioProject |
PRJNA417665 |
SRA |
SRP124611 |