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PMID: 25599178 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Integrated genome and transcriptome sequencing of the same cell.

Nature biotechnology ·Vol. 33 ·No. 3 ·2015-03-00 ·Pages 285-289

Dey SS, Kester L, Spanjaard B, Bienko M, van Oudenaarden A

Abstract

Single-cell genomics and single-cell transcriptomics have emerged as powerful tools to study the biology of single cells at a genome-wide scale. However, a major challenge is to sequence both genomic DNA and mRNA from the same cell, which would allow direct comparison of genomic variation and transcriptome heterogeneity. We describe a quasilinear amplification strategy to quantify genomic DNA and mRNA from the same cell without physically separating the nucleic acids before amplification. We show that the efficiency of our integrated approach is similar to existing methods for single-cell sequencing of either genomic DNA or mRNA. Further, we find that genes with high cell-to-cell variability in transcript numbers generally have lower genomic copy numbers, and vice versa, suggesting that copy number variations may drive variability in gene expression among individual cells. Applications of our integrated sequencing approach could range from gaining insights into cancer evolution and heterogeneity to understanding the transcriptional consequences of copy number variations in healthy and diseased tissues.

MeSH Terms
Cell Line, Tumor DNA/genetics Gene Expression Regulation Genome High-Throughput Nucleotide Sequencing/methods Humans RNA, Messenger/genetics,metabolism Single-Cell Analysis Transcriptome/genetics
Chemicals
RNA, Messenger DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Dey Siddharth S
Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Utrecht, The Netherlands. | University Medical Center Utrecht, Cancer Genomics Netherlands, Utrecht, The Netherlands.
Kester Lennart
Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Utrecht, The Netherlands. | University Medical Center Utrecht, Cancer Genomics Netherlands, Utrecht, The Netherlands.
Spanjaard Bastiaan
Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Utrecht, The Netherlands. | University Medical Center Utrecht, Cancer Genomics Netherlands, Utrecht, The Netherlands.
Bienko Magda
Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Utrecht, The Netherlands. | University Medical Center Utrecht, Cancer Genomics Netherlands, Utrecht, The Netherlands.
van Oudenaarden Alexander
Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Utrecht, The Netherlands. | University Medical Center Utrecht, Cancer Genomics Netherlands, Utrecht, The Netherlands.
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Article Info
Journal
Nature biotechnology
Abbr.
Nat Biotechnol
ISSN
1546-1696
Published
2015-03-00
Epub
2015-00-19
Pages
285-289
Language
English
Region
United States
NLM ID
9604648
PMCID
PMC4374170
Subset
IM
Grants
European Research Council · 294325 · International
Databases
GEO
Analysis Services
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