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Single-Cell Alternative Splicing Analysis with Expedition Reveals Splicing Dynamics during Neuron Differentiation.

Molecular cell · 2017 · Vol. 67 (1) · pp. 148-161.e5

Abstract

Alternative splicing (AS) generates isoform diversity for cellular identity and homeostasis in multicellular life. Although AS variation has been observed among single cells, little is known about the biological or evolutionary significance of such variation. We developed Expedition, a computational framework consisting of outrigger, a de novo splice graph transversal algorithm to detect AS; anchor, a Bayesian approach to assign modalities; and bonvoyage, a visualization tool using non-negative matrix factorization to display modality changes. Applying Expedition to single pluripotent stem cells undergoing neuronal differentiation, we discover that up to 20% of AS exons exhibit bimodality. Bimodal exons are flanked by more conserved intronic sequences harboring distinct cis-regulatory motifs, constitute much of cell-type-specific splicing, are highly dynamic during cellular transitions, preserve reading frame, and reveal intricacy of cell states invisible to conventional gene expression analysis. Systematic AS characterization in single cells redefines our understanding of AS complexity in cell biology.

Publication Types

["Journal Article"]

Keywords

MeSH Terms

["Algorithms", "Alternative Splicing", "Bayes Theorem", "Cell Line", "Computer Simulation", "Evolution, Molecular", "Gene Expression Regulation, Developmental", "Humans", "Kinetics", "Male", "Models, Genetic", "Nerve Tissue Proteins", "Neural Stem Cells", "Neurogenesis", "Neurons", "Phenotype", "Pluripotent Stem Cells", "RNA, Messenger", "Single-Cell Analysis"]

Funding

R01 HG004659 NHGRI NIH HHS (United States)
P30 NS047101 NINDS NIH HHS (United States)
U19 MH107367 NIMH NIH HHS (United States)
R01 AI123202 NIAID NIH HHS (United States)
R01 HD085902 NICHD NIH HHS (United States)
R01 NS075449 NINDS NIH HHS (United States)
R01 AI095277 NIAID NIH HHS (United States)

Linked Datasets (1)

GSE85908 GSE via ncbi_elink
GEO

Single-cell alternative splicing analysis with Expedition reveals splicing dynamics during neuron differentiation

Homo sapiens
428 data files
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P1_08_R1.fastq.gz RNA-Seq 612.1 MB
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P2_03_R1.fastq.gz RNA-Seq 1.3 GB
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P2_04_R1.fastq.gz RNA-Seq 2.3 GB
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P2_05_R1.fastq.gz RNA-Seq 1.3 GB
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P2_08_R1.fastq.gz RNA-Seq 947.8 MB
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P2_09_R1.fastq.gz RNA-Seq 1.3 GB
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P3_01_R1.fastq.gz RNA-Seq 6.3 GB
P3_02_R1.fastq.gz RNA-Seq 6.7 GB
P3_02_R1.fastq.gz RNA-Seq 6.7 GB
P3_03_R1.fastq.gz RNA-Seq 7.7 GB
P3_03_R1.fastq.gz RNA-Seq 7.7 GB
P4_01_R1.fastq.gz RNA-Seq 1.3 GB
P4_01_R1.fastq.gz RNA-Seq 1.3 GB
P4_06_R1.fastq.gz RNA-Seq 1.0 GB
P4_06_R1.fastq.gz RNA-Seq 1.0 GB
P4_07_R1.fastq.gz RNA-Seq 1.1 GB
P4_07_R1.fastq.gz RNA-Seq 1.1 GB
P4_08_R1.fastq.gz RNA-Seq 896.8 MB
P4_08_R1.fastq.gz RNA-Seq 896.8 MB
P4_09_R1.fastq.gz RNA-Seq 1.2 GB
P4_09_R1.fastq.gz RNA-Seq 1.2 GB
P4_10_R1.fastq.gz RNA-Seq 2.5 GB
P4_10_R1.fastq.gz RNA-Seq 2.5 GB
P4_11_R1.fastq.gz RNA-Seq 1.3 GB
P4_11_R1.fastq.gz RNA-Seq 1.3 GB
P4_12_R1.fastq.gz RNA-Seq 1.2 GB
P4_12_R1.fastq.gz RNA-Seq 1.2 GB
P6_01_R1.fastq.gz RNA-Seq 6.4 GB
P6_01_R1.fastq.gz RNA-Seq 6.4 GB
P6_02_R1.fastq.gz RNA-Seq 4.4 GB
P6_02_R1.fastq.gz RNA-Seq 4.4 GB
P6_03_R1.fastq.gz RNA-Seq 7.0 GB
P6_03_R1.fastq.gz RNA-Seq 7.0 GB
P6_04_R1.fastq.gz RNA-Seq 5.6 GB
P6_04_R1.fastq.gz RNA-Seq 5.6 GB
P7_01_R1.fastq.gz RNA-Seq 1.6 GB
P7_01_R1.fastq.gz RNA-Seq 1.6 GB
P7_02_R1.fastq.gz RNA-Seq 1.8 GB
P7_02_R1.fastq.gz RNA-Seq 1.8 GB
P7_03_R1.fastq.gz RNA-Seq 1.6 GB
P7_03_R1.fastq.gz RNA-Seq 1.6 GB
P7_04_R1.fastq.gz RNA-Seq 818.1 MB
P7_04_R1.fastq.gz RNA-Seq 818.1 MB
P7_05_R1.fastq.gz RNA-Seq 2.0 GB
P7_05_R1.fastq.gz RNA-Seq 2.0 GB
P7_06_R1.fastq.gz RNA-Seq 2.4 GB
P7_06_R1.fastq.gz RNA-Seq 2.4 GB
P7_07_R1.fastq.gz RNA-Seq 1.7 GB
P7_07_R1.fastq.gz RNA-Seq 1.7 GB
P7_08_R1.fastq.gz RNA-Seq 1.7 GB
P7_08_R1.fastq.gz RNA-Seq 1.7 GB
P7_09_R1.fastq.gz RNA-Seq 3.7 GB
P7_09_R1.fastq.gz RNA-Seq 3.7 GB
P7_10_R1.fastq.gz RNA-Seq 1.9 GB
P7_10_R1.fastq.gz RNA-Seq 1.9 GB
P7_11_R1.fastq.gz RNA-Seq 2.2 GB
P7_11_R1.fastq.gz RNA-Seq 2.2 GB
P7_12_R1.fastq.gz RNA-Seq 1.9 GB
P7_12_R1.fastq.gz RNA-Seq 1.9 GB
P8_01_R1.fastq.gz RNA-Seq 3.4 GB
P8_01_R1.fastq.gz RNA-Seq 3.4 GB
P8_02_R1.fastq.gz RNA-Seq 3.7 GB
P8_02_R1.fastq.gz RNA-Seq 3.7 GB
P8_05_R1.fastq.gz RNA-Seq 3.8 GB
P8_05_R1.fastq.gz RNA-Seq 3.8 GB
P8_06_R1.fastq.gz RNA-Seq 4.6 GB
P8_06_R1.fastq.gz RNA-Seq 4.6 GB
P8_10_R1.fastq.gz RNA-Seq 4.0 GB
P8_10_R1.fastq.gz RNA-Seq 4.0 GB
P8_11_R1.fastq.gz RNA-Seq 3.7 GB
P8_11_R1.fastq.gz RNA-Seq 3.7 GB
P9_01_R1.fastq.gz RNA-Seq 15.6 GB
P9_01_R1.fastq.gz RNA-Seq 15.6 GB
P9_02_R1.fastq.gz RNA-Seq 11.6 GB
P9_02_R1.fastq.gz RNA-Seq 11.6 GB
P9_03_R1.fastq.gz RNA-Seq 7.8 GB
P9_03_R1.fastq.gz RNA-Seq 7.8 GB
P9_04_R1.fastq.gz RNA-Seq 15.0 GB
P9_04_R1.fastq.gz RNA-Seq 15.0 GB

Analysis Pipelines (1)

geo_data_processing GSE85908