| Vector | lentivirus |
| Transgene/Inserted gene | CAR19 transgene |
| Regulatory element | human phosphoglycerate kinase (PGK) promoter |
| Vector production method | A self-inactivating (SIN) lentiviral vector stock for CAR19 transgene-coupled genome editing effects was generated for compliant T cell manufacture. The vector 3′ long terminal repeat (LTR) included multiplexed “minimal” U6 and H1 pol III promoter-sgRNA cassettes, which self-duplicated during reverse transcription and mediated editing effects at TRAC and CD52 target loci in the presence of Cas9. Healthy donor peripheral blood lymphocytes (PBLs) were activated and transduced using an automated CliniMacs Prodigy device, were returned to the chamber after off-device electroporation with Cas9 mRNA, and then were expanded before magnetic bead-mediated depletion of residual TCRαβ T cells. |
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