Preclinical Study CRISPR Target Regions CRISPR Delivery Method CAR T Product CAR T Target-Antigen CAR T delivery Method Overview Year [Ref]
Xenograft model of childhood ALL
TCR-β-chain
RNP Electroporation
TCR- CAR T-cells
CD19
Retroviral
TCR knockout CAR-T cells are a promising option for tertiary patients and showed no alloreactivity, despite their low persistency.
2020 (Stenger, Stief, Käuferle, et al., 2020)
Humanized murine model of glioma
TRAC, β2M, and PD-1
RNP Electroporation
Universal CAR-T cells resistant to PD-L1
EGFRvIII
AAV6 vector
CRISPR/Cas9 successfully generated universal CAR-T cells and enables them to resist against PD-L1. This CAR T-cell showed prolong survival and anti-tumor activity in mice.
2019 (Choi et al., 2019)
Humanized murine model of leukemia
TRAC
Cas9 mRNA Electroporation and gRNA lentiviral delivery
TCR- CAR T-cells
CD19
Lentiviral coupling CAR and gRNA
Separated delivery of CRISPR components may bring several advantages, including time-saving and low immunogenicity. TCR- CAR-T cells mediated highly effective leukemic eradication with less evidence of exhaustion
2018 (Georgiadis et al., 2018)
Orthotopic mouse glioma model
PD-1
Plasmid
PD1-deficient CD133 CAR T-cells
CD133
PiggyBac transposon system
PD1-deficient CD133 CAR-T cells showed enhanced cytotoxicity and proliferation but with cytokine secretion level. PD-1 destruction is associated with higher antitumor responses.
2018 (B. Hu et al., 2019)
Mice with pre-B acute lymphoblastic leukemia
TRAC
Cas9 mRNA and gRNA Electroporation
TCR- CAR T-cells
CD19
AAV6 vector
Knocking out of TRAC locus led to CAR T-cell potency and delayed effector CAR T differentiation and exhaustion.
2017 (Eyquem et al., 2017)
Xenograft tumor model of leukemia
PD-1
RNP Electroporation
PD1-deficient CD19 CAR T-cells
CD19
Lentiviral
This study shed light on PD-1 inhibitory roles in CAR T-cells. PD1-deficient CD19 CAR-T cells successfully eliminated tumor cells in vivo.
2017 (Rupp et al., 2017)
Mice with B cell precursor leukemia (Nalm6 or Nalm6-PDL1 tumor cells)
TCR, β2M, and PD1
Cas9 and gRNAs RNA Electroporation
Universal CAR-T cells resistant to PD-L1
PSCA and CD19
Lentiviral
TCR and β2M knockout CAR-T cells can be considered as universal CAR T-cells. and PD1 knockout led to enhanced in vivo anti-tumor activity. 2016 (Jiangtao Ren, Xiaojun Liu, et al., 2017)