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Single-Cell Immuno-Transcriptome Library
Description
Immune Repertoire (IR) refers to the diversity of antigen receptors on B and T lymphocytes, specifically the diversity of BCR (B-cell receptors) and TCR (T-cell receptors). BCRs and TCRs are typically composed of variable (V) regions, diversity (D) regions, joining (J) regions, and constant (C) regions. During genetic processes, the V(D)J regions undergo rearrangement or recombination, resulting in a wide variety of combinations and a rich diversity of BCR and TCR gene sequences. This collection of sequence diversity is known as the immune repertoire.
Single-Cell Immune Repertoire Sequencing (scVDJ-seq) utilizes the 10x Genomics platform's microfluidics and droplet encapsulation technologies to simultaneously detect transcriptome gene expression and TCR/BCR diversity at the single-cell level. This method not only provides insights into TCR/BCR clonotypes but also integrates transcriptome data to delve deeper into biological mechanisms. It offers a highly efficient platform for immuno-genomic research, with broad application prospects in fields such as tumor microenvironment studies, infectious diseases, and autoimmune diseases.
Workflow
Using the 10x Genomics platform, single cells are isolated and reverse transcribed into cDNA. The cDNA is then divided into two parts: one part is used for TCR/BCR V(D)J library construction, and the other part is used for 5' single-cell transcriptome library construction. Sequencing primers are added to both libraries. This process provides both 5' gene expression data and full-length V(D)J sequence information for each cell. The obtained data are then analyzed for cell subpopulation clustering, cell expression profiles, marker molecule identification, and immune repertoire clonotypes.
Advantages
- High Throughput: Detect immune repertoire diversity of tens of thousands of cells using the 10x Genomics single-cell capture platform.
- High Resolution: Obtain true single-cell level immune repertoire information.
- Comprehensive Information: Amplification covers the full-length V(D)J gene sequences for a more complete view of immune repertoire diversity.
- Double-Strand Accurate Matching: Efficiently match BCR heavy and light chains, as well as TCR α and β chains, at the single-cell level.
- Single-Cell Expression: Simultaneously measure immune repertoire (TCR/BCR) information and 5’ gene expression from the same cell.
- Extensive Experience: Over 100 tissue types and 3000+ single-cell samples prepared.
- Powerful Bioinformatics Analysis: Combine single-cell immune repertoire and gene expression analyses to uncover more cell interactions and provide personalized insights.
Applications
- Cancer Therapy: Investigate infiltrating lymphocytes in various types of cancer (such as colorectal cancer, ovarian cancer, liver cancer, melanoma, etc.), capture changes in the immune microenvironment during tumor development, and identify targets for immunotherapy to aid in more effective cancer treatment.
- Transplantation and Immune Reconstitution: During organ or bone marrow transplants, host rejection reactions can often lead to chronic graft-versus-host disease. Immune repertoire sequencing technology can assess the post-transplant T-cell repertoire to identify high-risk individuals for infections or relapse.
- Autoimmune Diseases: Analyze aberrantly expressed immune cells (T-cells or antibodies) to discover clinical diagnostic markers. Seek biomarkers superior to existing standards and study the mechanisms of the immune system in autoimmune conditions to provide a basis for disease diagnosis and treatment.
- Drug and Vaccine Evaluation: Assess peripheral blood samples after drug treatment to confirm if the drug induces an immune response and its efficacy. Obtain characteristic BCR (IG) sequences to shorten the antibody development process.
- Other Areas: Immune repertoire analysis provides crucial insights into infectious diseases, variations in immune profiles, and immunotherapy research.