本期文章：《细胞》：Online/在线发表

美国加州大学旧金山分校Justin Eyquem等研究人员合作发现，一种演化的AAV变体能够有效地对小鼠T细胞进行基因工程改造。2023年1月12日，《细胞》杂志在线发表了这项成果。

为了使鼠T细胞中有靶向的敲入，研究人员演化了ARK313，这是一种合成腺相关病毒（AAV），在鼠T细胞中表现出高转导效率。 研究人员进行了全基因组敲除筛查，并将QA2鉴定为ARK313感染的重要因素。 研究人员证明ARK313可用于无核能DNA递送，CRISPR-CAS9介导的敲入以及大型转基因的靶向整合。 ARK313可以在免疫能力模型中对Trac靶向的CAR-T和转基因TCR-T细胞进行临床前建模。 鼠T细胞中有效的基因靶向具有改善细胞疗法的巨大潜力，并在实验T细胞免疫学中开放了途径。

据悉，使用同源性修复将大型转基因的精确靶向对T细胞的靶向已成为过继细胞疗法和T细胞生物学的变化。 通过AAV传递DNA模板的效果极大地提高了敲入效率，但是当前AAV血清型的向性限制了它们对免疫缺陷小鼠模型中人类T细胞的使用。

附：英文原文

Title: An evolved AAV variant enables efficient genetic engineering of murine T cells

Author: William A. Nyberg, Jonathan Ark, Angela To, Sylvanie Clouden, Gabriella Reeder, Joseph J. Muldoon, Jing-Yi Chung, William H. Xie, Vincent Allain, Zachary Steinhart, Christopher Chang, Alexis Talbot, Sandy Kim, Alan Rosales, L. Patrick Havlik, Harold Pimentel, Aravind Asokan, Justin Eyquem

Issue&Volume: 2023-01-12

Abstract: 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.

DOI: 10.1016/j.cell.2022.12.022