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bioRxiv · 10.1101/2021.07.11.451925

Lentiviral vector optimization enhances the expression and cytotoxicity of chimeric antigen receptors

Guo, C.·Chen, H.·Yu, J.·Lu, H.·Guo, X.·Li, X.·Wang, T.·Zhi, L.·Niu, Z.·Zhu, W.
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Abstract

Adoptive chimeric antigen receptor (CAR)-modified T or NK cells (CAR-T/NK) have emerged as a novel form of disease treatment. Lentiviral vectors (LVs) are commonly employed to engineer T/NK cells for the efficient expression of CARs. This study reported for the first time the influence of single-promoter and dual-promoter LVs on the CAR expression and cytotoxicity of engineered NK cells. Our results demonstrated that the selected CAR exhibits both a higher expression level and a higher coexpression concordance with the GFP reporter in HEK-293T or NK92 cells by utilizing the optimized single-promoter pCDHsp rather than the original dual-promoter pCDHdp. After puromycin selection, the pCDHsp produces robust CAR expression and enhanced in vitro cytotoxicity of engineered NK cells. Therefore, infection with a single-promoter pCDHsp lentivector is recommended to prepare CAR-engineered cells. This research will help to optimize the production of CAR-NK cells and improve their functional activity, to provide CAR-NK cell products with better and more uniform quality.

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Guo, C., Chen, H., Yu, J., Lu, H., Guo, X., Li, X., Wang, T., Zhi, L., Niu, Z., Zhu, W.. 2021-07-12. Lentiviral vector optimization enhances the expression and cytotoxicity of chimeric antigen receptors. https://doi.org/10.1101/2021.07.11.451925

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