加州大学Justin Eyquem小组取得一项新突破。他们开发出了体内位点特异性工程重编程T细胞。该项研究成果发表在2026年3月18日出版的《自然》上。

在这里，课题组研究人员证明了稳定和细胞特异性的转基因表达可以通过大DNA有效载荷的体内位点特异性整合来实现。课题组开发了一种双载体系统来递送CRISPR-Cas9核糖核蛋白和DNA供体模板，分别对包膜递送载体和腺相关病毒主题进行主题化。课题组优化了这两种载体的T细胞特异性传递和基因靶向效率。通过将CAR转基因整合到T细胞特异性位点中，该研究组在B细胞发育不全、血液病和实体恶性肿瘤的人源化小鼠模型中产生了治疗水平的CAR - T细胞。这些发现为更有效、精确和广泛使用的T细胞疗法提供了一条途径。

据悉，工程T细胞，重新编程表达嵌合抗原受体（CAR）或T细胞受体（TCR），已经改变了癌症治疗，并正在探索作为自身免疫性和感染性疾病的治疗方法。通过基因组编辑来增强T细胞功能，无论是通过破坏内源性基因还是精确插入DNA有效载荷，都显示出相当大的前景。然而，体外制造过程漫长而昂贵，限制了这些疗法的可及性。在体内生成CAR - T细胞可以克服这些障碍，但目前的方法要么依赖于持久性有限的瞬时表达，要么依赖于缺乏特异性的DNA有效载荷的随机整合。

附：英文原文

Title: In vivo site-specific engineering to reprogram T cells

Author: Nyberg, William A., Bernard, Pierre-Louis, Ngo, Wayne, Wang, Charlotte H., Ark, Jonathan, Rothrock, Allison, Borgo, Gina M., Kimmerly, Gabriella R., Jung, Jae Hyung, Allain, Vincent, Hamilton, Jennifer R., Baldwin, Alisha, Stickels, Robert, Wyman, Sarah, Khan, Safwaan H., Lang, Shanshan, Marsh, Donna, Almudhfar, Niran, Novick, Catherine, Mortazavi, Yasaman, Zhang, Shimin, AbdElwakil, Mahmoud M., Sandoval, Luis R., Hwang, Sidney, Chu, Simon N., Jung, Hyuncheol, Liu, Chang, Sharma, Devesh, McCreary, Travis, Li, Zhongmei, Satpathy, Ansuman T., Carnevale, Julia, Rutishauser, Rachel L., Cromer, M. Kyle, Roybal, Kole T., Dodgson, Stacie E., Doudna, Jennifer A., Asokan, Aravind, Eyquem, Justin

Issue&Volume: 2026-03-18

Abstract: Engineered T cells, reprogrammed to express chimeric antigen receptors (CAR) or T cell receptors (TCR), have transformed cancer treatment and are being explored as therapeutics for autoimmune and infectious diseases. Enhancing T cell function through genome editing, either by disrupting endogenous genes or precisely inserting DNA payloads, has shown considerable promise1. However, the ex vivo manufacturing process is lengthy and costly, limiting accessibility of these therapies. In vivo generation of CAR T cells could overcome these barriers, but current methods rely either on transient expression with limited durability, or on random integration of DNA payloads that lack specificity. Here we demonstrate that stable and cell-specific transgene expression can be achieved through in vivo site-specific integration of large DNA payloads. We developed a two-vector system to deliver CRISPR–Cas9 ribonucleoproteins and a DNA donor template, using enveloped delivery vehicles and adeno-associated viruses, respectively. We optimized both vectors for T cell-specific delivery and gene-targeting efficiency. By integrating a CAR transgene into a T cell-specific locus, we generate therapeutic levels of CAR T cells in vivo in humanized mouse models of B cell aplasia, and haematological and solid malignancies. These findings offer a pathway to more efficient, precise and widely accessible T cell therapies.

DOI: 10.1038/s41586-026-10235-x

Source: https://www.nature.com/articles/s41586-026-10235-x