Figure 2.

Personalized adoptive T-cell therapy using gene editing. To perform personalized adoptive T-cell therapy, a biopsy of the patient's tumor can be subjected to whole-exome sequencing and RNA sequencing and identify the most immunogenic neoantigens using bioinformatic prediction algorithms. T cells specific for the predicted neoantigens can be isolated from the patient's tumor infiltrating lymphocytes (TILs) or peripheral blood mononuclear cells (PBMCs) using multimer peptide-MHC complexes (pMHC) libraries or dendritic cells transduced with neoantigen tandem minigenes, among other techniques. The two chains of the T-cell receptors (TCRs) can then be sequenced from single cells. Linear DNA encoding for these TCRs can be synthesized and electroporated together with gRNA and Cas9 into PBMCs from the same patient to generate neoantigen-specific T cells. These gene-edited T cells can be expanded and re-infused back into the patient. Multiplex knock-out can be used to increase the potency of T cells.