Related Experiment Video
Updated: Oct 20, 2025
Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
Published on: May 9, 2025
Bispecific binder redirected lentiviral vector enables in vivo engineering of CAR-T cells
Justin T Huckaby1, Elisa Landoni2, Timothy M Jacobs3
1UNC/NCSU Joint Department of Biomedical Engineering, UNC-Chapel Hill, Chapel Hill, North Carolina, USA.
Background:
Chimeric antigen receptor (CAR) T cells have shown considerable promise as a personalized cellular immunotherapy against B cell malignancies. However, the complex and lengthy manufacturing processes involved in generating CAR T cell products ex vivo result in substantial production time delays and high costs. Furthermore, ex vivo expansion of T cells promotes cell differentiation that reduces their in vivo replicative capacity and longevity.
Methods:
Here, to overcome these limitations, CAR-T cells are engineered directly in vivo by administering a lentivirus expressing a mutant Sindbis envelope, coupled with a bispecific antibody binder that redirects the virus to CD3+ human T cells.
Results:
This redirected lentiviral system offers exceptional specificity and efficiency; a single dose of the virus delivered to immunodeficient mice engrafted with human peripheral blood mononuclear cells generates CD19-specific CAR-T cells that markedly control the growth of an aggressive pre-established xenograft B cell tumor.
Conclusions:
These findings underscore in vivo engineering of CAR-T cells as a promising approach for personalized cancer immunotherapy.
