biostudies-literatureUnknown29Madison BBThe use of T cells from healthy donors for allogeneic chimeric antigen receptor T (CAR-T) cell cancer therapy is attractive because healthy donor T cells can produce versatile off-the-shelf CAR-T treatments. To maximize safety and durability of allogeneic products, the endogenous T cell receptor and major histocompatibility complex class I molecules are often removed via knockout of T cell receptor beta constant (<i>TRBC</i>) (or T cell receptor alpha constant [<i>TRAC</i>]) and <i>B2M</i>, respectively. However, gene editing tools (e.g., CRISPR-Cas9) can display poor fidelity, which may result in dangerous off-target mutations. Additionally, many gene editing technologies require T cell activation, resulting in a low percentage of desirable stem cell memory T cells (T_SCM). We cMolecular therapy. Nucleic acids979-995https://www.ebi.ac.uk/biostudies/studies/S-EPMC9481872biostudies-literatureCas-CLOVER is a novel high-fidelity nuclease for safe and robust generation of T_SCM-enriched allogeneic CAR-T cells.PMC9481872Marquez KOstertag EMPatil DTong MShedlock DJRichter MXi HLi XCoronella JWeiss LMadison BBTan YCranert SWang XMartin RfalseCas-CLOVER is a novel high-fidelity nuclease for safe and robust generation of T_SCM-enriched allogeneic CAR-T cells.The use of T cells from healthy donors for allogeneic chimeric antigen receptor T (CAR-T) cell cancer therapy is attractive because healthy donor T cells can produce versatile off-the-shelf CAR-T treatments. To maximize safety and durability of allogeneic products, the endogenous T cell receptor and major histocompatibility complex class I molecules are often removed via knockout of T cell receptor beta constant (<i>TRBC</i>) (or T cell receptor alpha constant [<i>TRAC</i>]) and <i>B2M</i>, respectively. However, gene editing tools (e.g., CRISPR-Cas9) can display poor fidelity, which may result in dangerous off-target mutations. Additionally, many gene editing technologies require T cell activation, resulting in a low percentage of desirable stem cell memory T cells (T_SCM). We c2022-01-01T00:00:00Z2022 Sep2025-04-22T03:30:22.507Z2025-04-05T20:45:39.18ZS-EPMC94818723618908010.1016/j.omtn.2022.06.003