Tailoring the resolution of single-cell RNA sequencing for primary cytotoxic T cells.

Details

Serval ID
serval:BIB_C79F07ADC7C7
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Tailoring the resolution of single-cell RNA sequencing for primary cytotoxic T cells.
Journal
Nature communications
Author(s)
Kanev K., Roelli P., Wu M., Wurmser C., Delorenzi M., Pfaffl M.W., Zehn D.
ISSN
2041-1723 (Electronic)
ISSN-L
2041-1723
Publication state
Published
Issued date
25/01/2021
Peer-reviewed
Oui
Volume
12
Number
1
Pages
569
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
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. Here, 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.
Keywords
Animals, Gene Expression Profiling/methods, High-Throughput Nucleotide Sequencing/methods, Humans, Mice, RNA/genetics, RNA, Messenger/genetics, Reproducibility of Results, Sequence Analysis, RNA/methods, Single-Cell Analysis/methods, T-Lymphocytes, Cytotoxic/cytology, T-Lymphocytes, Cytotoxic/metabolism
Pubmed
Open Access
Yes
Create date
23/02/2021 9:47
Last modification date
11/12/2021 7:38
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