Table S1 shows all lentiviral vectors used in this study, which were cloned according to our design by VectorBuilder (Chicago, IL, USA). Plasmid DNA was purified using a ZymoPURE II-EndoZero plasmid maxiprep kit (Zymo Research, Irvine, CA, USA). Unless otherwise specified, HEK293T cells in 125 cm2 flasks were transfected with Lipofectamine 3000 (Thermo Fisher Scientific) and DNA of lentiviral vector, second-generation packaging plasmid pCMV-dR8.2dvpr, and pCMV-VSVG at a 5:4:1 mass ratio. Where indicated, DNA of lentiviral vector, third-generation packaging plasmids pMDLg/pRRE and pRSV-Rev, plus pCMV-VSVG were used at a 5:2.5:1.25:1 mass ratio. dR8.2dvpr and pCMV-VSVG were gifts from Bob Weinberg via Addgene (Watertown, MA, USA), while pMDLg/pRRE and pRSV-Rev were gifts from Didier Trono via Addgene. Medium was exchanged after 6 h to Opti-MEM (Thermo Fisher Scientific) with 2 mM GlutaMAX and 5% FBS. Supernatants were collected 1 and 2 days later, then concentrated by precipitation with Lenti-X reagent (Takara Bio, Kusatsu, Japan) or by centrifugation at 18,600 × g relative centrifugal force (RCF) for 2 h at 4°C. Viral particles were resuspended in PBS or in X-VIVO 20 at 1/100 of the volume of the total supernatant. Viral titers were determined by serial dilution and transduction of HeLa cells in the presence of 8 μg/mL polybrene, and then flow cytometry for GFP expression 3 days later. Purification by Lenti-X produced higher titers (with HeLa cells) compared with ultracentrifugation. However, when matched to an equivalent MOI using the HeLa titration results, ultracentrifuged viral preparations resulted in greater percentages of transduced NK cells (data not shown).

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