An evolved AAV variant enables efficient genetic engineering of murine T cells.
Journal Article (Journal Article)
Precise targeting of large transgenes to T cells using homology-directed repair has been transformative for adoptive cell therapies and T cell biology. Delivery of DNA templates via adeno-associated virus (AAV) has greatly improved knockin efficiencies, but the tropism of current AAV serotypes restricts their use to human T cells employed in immunodeficient mouse models. To enable targeted knockins in murine T cells, we evolved Ark313, a synthetic AAV that exhibits high transduction efficiency in murine T cells. We performed a genome-wide knockout screen and identified QA2 as an essential factor for Ark313 infection. We demonstrate that Ark313 can be used for nucleofection-free DNA delivery, CRISPR-Cas9-mediated knockouts, and targeted integration of large transgenes. Ark313 enables preclinical modeling of Trac-targeted CAR-T and transgenic TCR-T cells in immunocompetent models. Efficient gene targeting in murine T cells holds great potential for improved cell therapies and opens avenues in experimental T cell immunology.
- Nyberg, WA; Ark, J; To, A; Clouden, S; Reeder, G; Muldoon, JJ; Chung, J-Y; Xie, WH; Allain, V; Steinhart, Z; Chang, C; Talbot, A; Kim, S; Rosales, A; Havlik, LP; Pimentel, H; Asokan, A; Eyquem, J
- January 19, 2023
Volume / Issue
- 186 / 2
Start / End Page
- 446 - 460.e19
Electronic International Standard Serial Number (EISSN)
Digital Object Identifier (DOI)
- United States