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| Status |
Public on Dec 13, 2020 |
| Title |
Tailoring the resolution of single-cell RNA sequencing for primary cytotoxic T cells |
| Organisms |
Homo sapiens; Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Single-cell RNA sequencing in principle offers unique opportunities to improve the efficacy of contemporary T-cell based immunotherapy against cancer. The use of high-quality single-cell data will aid our incomplete understanding of molecular programs determining the differentiation and functional heterogeneity of cytotoxic T lymphocytes (CTLs) allowing for optimal therapeutic design. So far a major obstacle to high depth single-cell analysis of CTLs is the minute amount of RNA available leading to low capturing efficacy. To overcome this we tailor a droplet-based approach for high-throughput analysis (tDrop-seq) and a plate-based method for high-performance in-depth CTL analysis (tSCRB-seq). The latter gives on average a 15-fold higher number of captured transcripts per gene compared to droplet-based technologies. The improved dynamic range of gene detection gives tSCRB-seq an edge in resolution sensitive downstream applications such as graded high confidence gene expression measurements and cluster characterization. We demonstrate the power of tSCRB-seq by revealing the subpopulation-specific expression of co-inhibitory and co-stimulatory receptor targets of key importance for immunotherapy.
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| Overall design |
This study groups a total of 27 samples of single-cell RNA sequencing with two different protocols (Drop-seq and SCRB-seq) and their modifications.
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| Contributor(s) |
Kanev K, Roelli P, Wu M, Wurmser C, Delorenzi M, Pfaffl MW, Zehn D |
| Citation(s) |
33495472 |
| Submission date |
Dec 12, 2020 |
| Last update date |
Feb 02, 2021 |
| Contact name |
Patrick Roelli |
| E-mail(s) |
patrick.roelli@gmail.com
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| Organization name |
TUM
|
| Department |
Tierphysiologie & Immunologie
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| Lab |
Dr. Zehn
|
| Street address |
Weihenstephaner Berg 3
|
| City |
Freising |
| ZIP/Postal code |
85354 |
| Country |
Germany |
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| Platforms (2) |
| GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
| GPL22245 |
Illumina HiSeq 2500 (Homo sapiens; Mus musculus) |
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| Samples (27)
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| Relations |
| BioProject |
PRJNA684762 |
| SRA |
SRP297763 |
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