From: Chimeric antigen receptor natural killer (CAR-NK) cell design and engineering for cancer therapy
Method | Advantages | Disadvantages | GMP compliance |
---|---|---|---|
Lentivirus | High efficiency | Potential genotoxicity owing to LTR sequences | Third generation is considered safe enough for clinical use |
Favorable safety profile | Difficult to obtain high titer LV | ||
Transduction of resting NK | |||
Retrovirus | Long history in clinical trials | Risk of insertional oncogenesis | Compatible; successfully used to generate CAR-NK for clinical trials |
Relatively easy to produce | Transgene capacity is lower than LV | ||
Requires actively dividing cells | |||
mRNA electroporation | Cheaper than viral production | Transient expression | Compatible |
No integration risks | |||
High transduction rates | |||
Transposon | Transgene capacity higher than for viruses | Risk of insertion into genome | Compatible |
Cheap, easy to produce | |||
CRISPR/Cas9 | Precise gene modification | Complicated to design the HDR template | Not yet applied in CAR-NK clinical trials |
Ability to use endogenous promoter of choice | Efficiency is hard to control |