Hamberger, M.; Neuhoff, M.-T.; Torres, C. M.; Tan, C. L.; Boschert, T.; Errerd, A.; Lindner, J. M.; Platten, M.; Green, E. W.

Personalised cell therapies utilising T cell receptors (TCRs) show great promise, but synthesising and testing TCRs lags behind our ability to sequence TCR repertoires. Current costs of TCR synthesis limit our ability to generate sufficiently large datasets to train the next generation of machine learning models to determine TCR targets from TCR sequence alone. We built a modular Golden Gate mediated TCR cloning system that enables rapid, single-step manufacture of TCR sequences into diverse expression vectors with free choice of engineered and native TCR constant regions. We provide modules for cloning human and mouse {beta} and {gamma}{delta} TCRs, and show how implementing cell-free manufacturing facilitates propagation of modules, generates testable TCRs within 24 hours, and scales to making 1000s of TCRs for under $4 each. makeTCR is available as an free, open-source, extensible, graphical platform to simplify, standardise, cheapen and accelerate TCR functionality testing for personalised medicine and beyond.