By integrating scRNA-seq and TCR repertoire analysis, gene expression information and TCRα/β pair sequences necessary for clone reconstitution can be simultaneously analyzed for individual T cells. Based on the characteristics and frequency of each clone, it is possible to narrow down clones that are promising for TCR gene therapy and other applications.
* For TCR repertoireanalysis, please also refer to the TCR repertoire analysis page. Here, TCR repertoireanalysis is performed at the single cell level.
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Single-cell RNA-seq is available with either whole transcriptome amplification (WTA) or targeted sequencing using BD's Immune Response Panel.
*Target sequencing can optionally analyze genes not included in the target panel.
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Capable of up to 20,000-30,000 cells/analysis, pairing efficiency of mouse and human bio-derived TCRα/β is around 20%-80% (depending on cell activation state).
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Simultaneous analysis with protein using BD's Abseq, BioLegend's TotalSeq, etc. is also available
TAS-Seq2 detects about 3 times more genes than BD's WTA kit
TAS-Seq2 has a higher T-cell receptor pairing rate than BD's WTA kit
Example of analysis of tumor-infiltrating CD8⁺T cells in a B16F10 subcutaneously implanted tumor mouse model upon anti-PD-L1 treatment (WTA)
Approx. 50,000 reads/cell
EXPRESSION PATTERNS OF REPRESENTATIVE CD8+ T CELL MARKER GENES
BD WTA
Clusters identified by TAS-Seq2
0: terminally exhausted, 1: effector, 2: proliferated, 3: exhausted progenitor-1, 4: exhausted progenitor-2, 5: naïve-like, 6: stem-like, 7: dying cell
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TAS-Seq2 can identify CD8+ T cell subsets that are difficult to identify with BD's WTA kit
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TAS-Seq2 can clearly detect genes involved in the regulation of CD8+ T cell function with a lower drop-out rate
Reverse transcription
5'末端へのへのPolyA付加
Self-hybridization and elongation
TCR Gene Amplification
TCR a/b library
Matters to be checked in advance
* For multiplex analysis, please refer to the FAQ page "About Multiplex Analysis".
How to request
Sample acceptance
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cDNA synthesized by our specified protocol (IGT will undertake cDNA amplification and subsequent steps)
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Pre-fractionated frozen T cell specimens (cDNA synthesis and beyond contracted by IGT)
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Unfractionated frozen cell specimens (IGT is entrusted with t cell preparation and beyond)
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Cryopreserved tissue specimens (IGT is entrusted with the preparation of cell suspensions and beyond)
* When cryopreserving cells and tissues, please use the preservation solution and freezing method specified by us.
How to send samples
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Please make sure that the sample is dry and leak-free, and send it to the shipping address below. (Available until 5:00 p.m. on weekdays)
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If shipping frozen tissue or cells, please include enough dry ice to maintain the frozen state and ship frozen.
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Please refer to "Sample Shipping Address" for shipping address.
* *Except for remote areas, please send by next morning (except Saturdays, Sundays, and national holidays).
Send samples to
1F Building 17, Tokyo University of Science, Noda Campus
2669 Yamazaki, Noda, Chiba 278-0022, Japan
ImmunoGeneTeqs, Inc.
Lead Time
From sample receipt
2.5 months
Deliverables
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Work report
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Complete sequence data
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Mapping result files, etc.
The official report will be delivered on a hard drive at the time of delivery
A mapping analysis report will be included in the formal report
An example of the report can be seen in the example of the analysis of mouse tumor-infiltrating T cells
scTCR-seq (WTA)
Reference price
1
2
3
4
scTCR-seq(targeted RNA-seq)
Reference price
1
2
3
4
Example of analysis of mouse tumor-infiltrating T cells
Tumor-infiltrating T cells (14,100 cells) were classified into 10 clusters and visualized by dimensional compression; clones sharing the TCR were then defined and the properties of each clone were evaluated; data consisted of single cell RNA-seq data and single cell TCR repertoireanalysis data.
The actual report will also be in this style
Authors：Hiroyasu Aoki, Masahiro Kitabatake, Haruka Abe, Peng Xu, Mikiya Tsunoda, Shigeyuki Shichino, Atsushi Hara, Noriko Ouji-Sageshima, Chihiro Motozono, Toshihiro Ito, Kouji Matsushima, and Satoshi Ueha
Journal, 掲載日：Cell Reports, 2024年3月8日
