CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification
High-throughput sequencing has allowed for unprecedented detail in gene expression analyses, yet its efficient application to single cells is challenged by the small starting amounts of RNA. We have developed CEL-Seq, a method for overcoming this limitation by barcoding and pooling samples before li...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2012-09-01
|
| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124712002288 |
| _version_ | 1811278805739765760 |
|---|---|
| author | Tamar Hashimshony Florian Wagner Noa Sher Itai Yanai |
| author_facet | Tamar Hashimshony Florian Wagner Noa Sher Itai Yanai |
| author_sort | Tamar Hashimshony |
| collection | DOAJ |
| description | High-throughput sequencing has allowed for unprecedented detail in gene expression analyses, yet its efficient application to single cells is challenged by the small starting amounts of RNA. We have developed CEL-Seq, a method for overcoming this limitation by barcoding and pooling samples before linearly amplifying mRNA with the use of one round of in vitro transcription. We show that CEL-Seq gives more reproducible, linear, and sensitive results than a PCR-based amplification method. We demonstrate the power of this method by studying early C. elegans embryonic development at single-cell resolution. Differential distribution of transcripts between sister cells is seen as early as the two-cell stage embryo, and zygotic expression in the somatic cell lineages is enriched for transcription factors. The robust transcriptome quantifications enabled by CEL-Seq will be useful for transcriptomic analyses of complex tissues containing populations of diverse cell types. |
| first_indexed | 2024-04-13T00:42:10Z |
| format | Article |
| id | doaj.art-7a5c4be316904d24850737cadd97a01b |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-04-13T00:42:10Z |
| publishDate | 2012-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-7a5c4be316904d24850737cadd97a01b2022-12-22T03:10:06ZengElsevierCell Reports2211-12472012-09-012366667310.1016/j.celrep.2012.08.003CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear AmplificationTamar Hashimshony0Florian Wagner1Noa Sher2Itai Yanai3Department of Biology, Technion-Israel Institute of Technology, Haifa 32000, IsraelDepartment of Biology, Technion-Israel Institute of Technology, Haifa 32000, IsraelDepartment of Biology, Technion-Israel Institute of Technology, Haifa 32000, IsraelDepartment of Biology, Technion-Israel Institute of Technology, Haifa 32000, IsraelHigh-throughput sequencing has allowed for unprecedented detail in gene expression analyses, yet its efficient application to single cells is challenged by the small starting amounts of RNA. We have developed CEL-Seq, a method for overcoming this limitation by barcoding and pooling samples before linearly amplifying mRNA with the use of one round of in vitro transcription. We show that CEL-Seq gives more reproducible, linear, and sensitive results than a PCR-based amplification method. We demonstrate the power of this method by studying early C. elegans embryonic development at single-cell resolution. Differential distribution of transcripts between sister cells is seen as early as the two-cell stage embryo, and zygotic expression in the somatic cell lineages is enriched for transcription factors. The robust transcriptome quantifications enabled by CEL-Seq will be useful for transcriptomic analyses of complex tissues containing populations of diverse cell types.http://www.sciencedirect.com/science/article/pii/S2211124712002288 |
| spellingShingle | Tamar Hashimshony Florian Wagner Noa Sher Itai Yanai CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification Cell Reports |
| title | CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification |
| title_full | CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification |
| title_fullStr | CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification |
| title_full_unstemmed | CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification |
| title_short | CEL-Seq: Single-Cell RNA-Seq by Multiplexed Linear Amplification |
| title_sort | cel seq single cell rna seq by multiplexed linear amplification |
| url | http://www.sciencedirect.com/science/article/pii/S2211124712002288 |
| work_keys_str_mv | AT tamarhashimshony celseqsinglecellrnaseqbymultiplexedlinearamplification AT florianwagner celseqsinglecellrnaseqbymultiplexedlinearamplification AT noasher celseqsinglecellrnaseqbymultiplexedlinearamplification AT itaiyanai celseqsinglecellrnaseqbymultiplexedlinearamplification |