A preparation of autologous T-lymphocytes that are non-virally gene-edited with the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 nuclease complex to integrate a chimeric antigen receptor (CAR) targeting the tumor-associated antigen (TAA) CD19 in the locus of the programmed cell death 1 (PD-1; PDCD1; CD279; programmed death-1) gene, with potential immunomodulating and antineoplastic activities. Upon administration, autologous PD1-knockout CD19-specific CAR T-cells BRL-201 specifically target and bind to CD19-expressing tumor cells, thereby selectively lysing CD19-expressing tumor cells. CD19 antigen is a B-cell specific cell surface antigen overexpressed in B-cell lineage malignancies. Expression of PD-1, an inhibitory receptor expressed on activated T-cells, plays a key role in cytotoxic T-lymphocyte (CTL) suppression, T-cell exhaustion and CTL apoptosis. PD-1 knockout may abrogate T-cell exhaustion and increase T-cell activity and cytotoxicity.