In vivo engineering of murine T cells using the evolved adeno-associated virus variant Ark313
Genetic engineering of T cells in mouse models is essential for investigating immune mechanisms. We aimed to develop an approach to manipulate T cells in vivo using an evolved adeno-associated virus (AAV) capsid named Ark313. Delivery of a transient transgene expression cassette was feasible using Ark313, and this serotype outperformed natural serotypes. A single intravenous injection of a Cre recombinase-expressing Ark313 in the Ai9 fluorescent reporter mouse model achieved permanent genetic modifications of T cells. Ark313 facilitated in vivo gene editing in both tissue-resident and splenic T cells and validation of immunotherapy targets in solid tumor models. Ark313 delivered large DNA donor templates to T cells in vivo and integrated transgenes in primary CD4<sup>+</sup> and CD8<sup>+</sup> T cells, including naive T cells. Ark313-mediated transgene delivery presents an efficient approach to target mouse T cells in vivo and a resource for the interrogation of T cell biology and for immunotherapy applications.
Authors
- Chun Jimmie Ye (ORCID: https://orcid.org/0000-0001-6560-3783)
- Sofia Caryotakis (ORCID: https://orcid.org/0000-0003-3522-0022)
- Kelly Kersten (ORCID: https://orcid.org/0000-0001-7615-2594)
- Joseph J. Muldoon (ORCID: https://orcid.org/0000-0003-3784-3565)
- Aravind Asokan (ORCID: https://orcid.org/0000-0001-5563-4877)
- Nicholas F. Kuhn (ORCID: https://orcid.org/0000-0001-5634-0846)
- Jonathan Ark (ORCID: https://orcid.org/0000-0003-4991-1422)
- Mingcheng Li (ORCID: https://orcid.org/0000-0003-2640-4305)
- Charlotte H. Wang (ORCID: https://orcid.org/0000-0001-7588-2077)
- P Bernard (ORCID: https://orcid.org/0000-0001-6567-619X)
- Peixin Amy Chen (ORCID: https://orcid.org/0000-0002-5946-1632)
- Justin Eyquem (ORCID: https://orcid.org/0000-0001-8262-1190)
- Sylvanie Clouden
- Maya Lopez-Ichikawa (ORCID: https://orcid.org/0000-0001-8609-1561)
- James M. Gardner (ORCID: https://orcid.org/0000-0003-3470-9520)
- Jin Seo
- Alan Rosales (ORCID: https://orcid.org/0000-0001-6986-2985)
- Gabriella R. Kimmerly (ORCID: https://orcid.org/0000-0002-2920-2218)
- Ari B. Molofsky (ORCID: https://orcid.org/0000-0003-0764-3175)
- Celeste Garza
- Roberto R. Ricardo-González (ORCID: https://orcid.org/0000-0002-6135-4035)
- Zhongmei Li (ORCID: https://orcid.org/0000-0001-8616-9568)
- Anita Qualls (ORCID: https://orcid.org/0000-0003-0576-8863)
- William A. Nyberg (ORCID: https://orcid.org/0000-0002-9982-1305)
- Hong-Erh Liang (ORCID: https://orcid.org/0000-0003-0620-7224)
- Chang Liu (ORCID: https://orcid.org/0000-0001-8189-5667)
- E. Wells
- Katherine Simon (ORCID: https://orcid.org/0000-0001-9232-2653)
Institutions
- Gladstone Institutes (US)
- Duke University (US)
- University of California, San Francisco (US)
Publication Details
- Journal
- Immunity
- Published
- 2025-02-01
- DOI
- https://doi.org/10.1016/j.immuni.2025.01.009
- Citations
- 18
- Primary Topic
- Virus-based gene therapy research
- Type
- article
- Field-Weighted Citation Impact
- 32.71
Funders
- Cancer Research Institute
- Parker Institute for Cancer Immunotherapy
- Svenska Sällskapet för Medicinsk Forskning
- National Institutes of Health