ArticlePloS one2011
A practical approach to T-cell receptor cloning and expression.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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35 citing papers in PubMed, 53 citations in OpenAlex.
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- Efficient CAR T cell targeting of the CA125 extracellular repeat domain of MUC16.Journal for immunotherapy of cancer · 2024Article
- CD20 expression regulates CD37 levels in B-cell lymphoma - implications for immunotherapies.Oncoimmunology · 2024Article
- ALPL-1 is a target for chimeric antigen receptor therapy in osteosarcoma.Nature communications · 2023Article
- TCR sequencing and cloning methods for repertoire analysis and isolation of tumor-reactive TCRs.Cell reports methods · 2023Article
- Targeting the melanoma-associated antigen CSPG4 with HLA-C*07:01-restricted T-cell receptors.Frontiers in immunology · 2023Article
- Mechanism of CD79A and CD79B Support for IgM+ B Cell Fitness through B Cell Receptor Surface Expression.Journal of immunology (Baltimore, Md. : 1950) · 2022Article
- Targeting Telomerase with an HLA Class II-Restricted TCR for Cancer Immunotherapy.Molecular therapy : the journal of the American Society of Gene Therapy · 2021Article
- Competition-Based Cell Assay Employing Soluble T Cell Receptors to Assess MHC Class II Antigen Processing and Presentation.The AAPS journal · 2021Article
- Targeting KRAS mutations with HLA class II-restricted TCRs for the treatment of solid tumors.Oncoimmunology · 2021Article
- Colorectal cysts as a validating tool for CAR therapy.BMC biotechnology · 2020Article
- A bicistronic vector backbone for rapid seamless cloning and chimerization of αβT-cell receptor sequences.PloS one · 2020Article
- Preclinical development of CD37CAR T-cell therapy for treatment of B-cell lymphoma.Blood advances · 2019Article
- Chimeric antigen receptor preparation from hybridoma to T-cell expression.Antibody therapeutics · 2019Article
- TIGIT and PD-1 Mark Intratumoral T Cells with Reduced Effector Function in B-cell Non-Hodgkin Lymphoma.Cancer immunology research · 2019Article
- NK cells specifically TCR-dressed to kill cancer cells.EBioMedicine · 2019Article
- Identification of α-fetoprotein-specific T-cell receptors for hepatocellular carcinoma immunotherapy.Hepatology (Baltimore, Md.) · 2018Article
- Human c-SRC kinase (CSK) overexpression makes T cells dummy.Cancer immunology, immunotherapy : CII · 2018Article
- Artesunate shows potent anti-tumor activity in B-cell lymphoma.Journal of hematology & oncology · 2018Article
- A TCR-based Chimeric Antigen Receptor.Scientific reports · 2017Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Although cloning and expression of T-cell Receptors (TcRs) has been performed for almost two decades, these procedures are still challenging. For example, the use of T-cell clones that have undergone limited expansion as starting material to limit the loss of interesting TcRs, must be weighed against the introduction of mutations by excess PCR cycles. The recent interest in using specific TcRs for cancer immunotherapy has, however, increased the demand for practical and robust methods to rapidly clone and express TcRs. Two main technologies for TcR cloning have emerged; the use of a set of primers specifically annealing to all known TcR variable domains, and 5'-RACE amplification. We here present an improved 5'-RACE protocol that represents a fast and reliable way to identify a TcR from 10(5) cells only, making TcR cloning feasible without a priori knowledge of the variable domain sequence. We further present a detailed procedure for the subcloning of TcRα and β chains into an expression system. We show that a recombination-based cloning protocol facilitates simple and rapid transfer of the TcR transgene into different expression systems. The presented comprehensive method can be performed in any laboratory with standard equipment and with a limited amount of starting material. We finally exemplify the straightforwardness and reliability of our procedure by cloning and expressing several MART-1-specific TcRs and demonstrating their functionality.
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