Leukopaks from deidentified healthy donors with Institutional Review Board-approved consent forms and protocols were purchased from StemCell Technologies (200-0092). Residuals from leukoreduction chambers after Trima Apheresis from deidentified healthy donors with Institutional Review Board-approved consent forms and protocols were purchased from Vitalant. All of the mice in this study were treated following a protocol (AN182757) approved by the UCSF Institutional Animal Care and Use Committee (IACUC). Plasmids For transient GFP expression, a scAAV plasmid with a CMV enhancer chicken B-actin intron (CAG) promoter was used (scAAV-CAG-GFP, Addgene, 83279). A similar ssAAV (ssAAV-CAG-GFP) has been used for biodistribution studies (Addgene, 28014). A plasmid encoding the HIV protein gag fused to Cas9 expressing four Nuclear Localization Signal (4×NLS) was used to package Cas9 into EDVs59. For integrating genes at the TRAC locus, we used the same homology arms and sgRNA sequence as previously described1. An EGFRt sequence was cloned in following a 1928z-1XX sequence33. To potentially improve nuclease activity when combined with a Cas9-containing EDV, a U6 promoter expressing the sgRNA for targeting TRAC was cloned into the plasmids upstream of the left homology arm (pAAV-U6/TRAC-TRAC-1928-1XX-P2A-EGFRt). The extracellular domain of a BCMA-targeting CAR was used to replace the CD19-binding sequence to generate...

Human peripheral blood Leukopaks enriched for peripheral blood mononuclear cells (PBMCs) from deidentified healthy donors were purchased from StemCell Technologies (200-0092). CD3+ T cells were isolated using the EasySep Human T cell isolation kit (100-069) per the manufacturer-provided instructions. After...

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BioIVT Introduces VivoSTART™ Cryopreserved GMP Leukopaks for Improved Cell Therapy Development  BioSpace