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.

Full Text

Duke Authors

Cited Authors

  • 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

Published Date

  • January 19, 2023

Published In

Volume / Issue

  • 186 / 2

Start / End Page

  • 446 - 460.e19

PubMed ID

  • 36638795

Electronic International Standard Serial Number (EISSN)

  • 1097-4172

Digital Object Identifier (DOI)

  • 10.1016/j.cell.2022.12.022


  • eng

Conference Location

  • United States