Refactored intracellular domains enhance CAR expression and function
- 1Generate Biomedicines
Abstract
Chimeric antigen receptor (CAR) intracellular domains (ICDs) couple antigen recognition to T-cell activation. Here we worked with AI agents to refactor CAR ICDs, proposing designs and refining them as measurements accumulated over three rounds. This approach yielded KRGR-{zeta}1-40, a 45-amino-acid ICD less than a third the length of the clinical 4-1BB-CD3{zeta} (BB{zeta}) ICD. Compared with BB{zeta}, KRGR-{zeta}1-40 increased CAR surface expression and antigen-driven IFN-{gamma} release, without detectable antigen-independent IFN-{gamma} release. KRGR-{zeta}1-40 extended CAR expression and target-cell killing after transient mRNA delivery, supported the expression of four CARs from a shared mRNA dose, and enhanced B-cell depletion after lentiviral and lipid-nanoparticle delivery. These results show that iterative agent-guided design with experimental feedback can yield compact ICDs that enhance CAR expression and sustain CAR T-cell function.
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BibTeX
@misc{bai2026refactored,
title = {{Refactored intracellular domains enhance CAR expression and function}},
author = {L. Bai and A. Kumar and H. J. Desai and J. Li and D. P. de Alwis and D. Hazuda and G. Grigoryan and W. Wang and J. Yi},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.28.762481},
url = {https://www.biorxiv.org/content/10.64898/2026.10.28.762481v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Refactored intracellular domains enhance CAR expression and function
AU - L. Bai
AU - A. Kumar
AU - H. J. Desai
AU - J. Li
AU - D. P. de Alwis
AU - D. Hazuda
AU - G. Grigoryan
AU - W. Wang
AU - J. Yi
PY - 2026
DA - 2026/10/28
PB - bioRxiv
DO - 10.64898/2026.10.28.762481
UR - https://www.biorxiv.org/content/10.64898/2026.10.28.762481v1
AB - Chimeric antigen receptor (CAR) intracellular domains (ICDs) couple antigen recognition to T-cell activation. Here we worked with AI agents to refactor CAR ICDs, proposing designs and refining them as measurements accumulated over three rounds. This approach yielded KRGR-{zeta}1-40, a 45-amino-acid ICD less than a third the length of the clinical 4-1BB-CD3{zeta} (BB{zeta}) ICD. Compared with BB{zeta}, KRGR-{zeta}1-40 increased CAR surface expression and antigen-driven IFN-{gamma} release, without detectable antigen-independent IFN-{gamma} release. KRGR-{zeta}1-40 extended CAR expression and target-cell killing after transient mRNA delivery, supported the expression of four CARs from a shared mRNA dose, and enhanced B-cell depletion after lentiviral and lipid-nanoparticle delivery. These results show that iterative agent-guided design with experimental feedback can yield compact ICDs that enhance CAR expression and sustain CAR T-cell function.
ER -