Skip to main content

Rapid Cloning of T Cell Receptors (TCRs) (SJ-16-0001)/ (Rapid cloning, expression, and functional characterization of paired αβ and γδ TCR chains from single cell analysis)

St. Jude Reference #SJ-16-0001


Transgenic expression of antigen-specific T cell receptor (TCR) genes is a promising approach for immunotherapy against infectious diseases and cancers. A key to the efficient application of this approach is the rapid and specific isolation and cloning of TCRs. Researchers at St. Jude have developed a novel method to rapidly clone, express and characterize the function of paired αβ and γδ TCR chains from single cells. The platform addresses the non-specific, labor-intensive, and time-consuming issues of traditional PCR-based cloning and it provides a relatively high-throughput, accurate, and efficient method of TCR engineering for therapeutic or research applications.

The researchers demonstrated the capability of cloning influenza-specific TCRs within 10 days using single cell PCR and Gibson Assembly techniques. This process can be accelerated to 5 days by generating receptor libraries, requiring only the exchange of the antigen-specific CDR3 region into an existing backbone. The functional activity of these TCRs can be characterized in a novel reporter cell line for screening of TCR specificity and avidity.

By generating a library of specific TCR constructs reactive against a range of viruses and HLA types, TCR-directed therapies could be used prophylactically or immediately at the earliest signs of viral reactivation or to target conserved or patient-specific tumor antigens.

In addition to therapeutic applications, the protocol significantly improves the workflow for cloning and expressing TCRs for study in vitro. 



Paired T-cell receptors (TCRs), TCR screening, stem cell transplant


Granted Patents or Published Applications

U. S. Patent application submitted, published international application WO 2017/096239


Related Scientific References

Xi-zhi J Guo, Pradyot Dash, Matthew Calverley, Suzanne Tomchuck, Mari H Dallas, and Paul G Thomas, Rapid cloning, expression, and functional characterization of paired αβ and γδ T-cell receptor chains from single-cell analysis. Mol Ther Methods Clin Dev. 2016; 3: 15054. Published online 2016 Jan 27.

Wang, G.C., Dash, P., McCullers, J.A., Doherty, P.C., and Thomas, P.G., T cell receptor αβ diversity inversely correlates with pathogen-specific antibody levels in human cytomegalovirus infection. Sci Transl Med 4, 128ra42 (2012).

Han, A et al., Linking T-cell receptor sequence to functional phenotype at the single cell level. Nature Biotech. 32(7): 684-692 (epub June 22, 2014).  

Licensing Opportunities

This invention enables a reduction in cost, time and reagents can get the cost per cell to under $1 per cell, compared with the standard sequencing technique which is now typically $6 per cell, it may be able to be used for cytomegalovirus (CMV) or other herpesvirus-positive patients and cancer patients in patient specific therapy, or it may be used as a molecular tool (i.e. metrics of immune health; better indicator of potent immune status) for drug development. If you are interested in licensing this technology in one of these or any other fields, please contact

Related Links


Contact the Office of Technology Licensing (Phone: 901-595-2342, Fax: 901-595-3148) for more information.