美国格莱斯顿-加州大学旧金山分校基因组免疫学研究所Luke A. Gilbert小组的最新研究提出了人类原代T细胞的综合表观遗传和遗传编程。该项研究成果发表在2025年10月21日出版的《自然—生物技术》上。

该研究团队开发了一个全RNA平台，用于在原代人T细胞中高效、持久和多路的表观遗传编程，稳定地关闭或打开内源性基因，为CRISPRoff和CRISPRon表观遗传编辑。课题组实现了多种靶向基因组元件的表观遗传编程，而不需要维持CRISPR系统的表达。CRISPRoff介导的基因沉默通过大量细胞分裂、T细胞刺激和体内过继转移来维持，避免了多重Cas9介导的基因组编辑所固有的细胞毒性或染色体异常。最后，该研究组成功地将基因和表观遗传工程主题正交CRISPR Cas12a-dCas9系统结合起来，靶向嵌合抗原受体（CAR）敲入和CRISPRoff沉默治疗相关基因，以改善临床前CAR- t细胞介导的体内肿瘤控制和生存。

据了解，细胞治疗中基因表达的靶向表观遗传工程将允许编程所需的表型，而不会出现与基于双链断裂的基因编辑方法相关的许多挑战和安全风险。

附：英文原文

Title: Integrated epigenetic and genetic programming of primary human T cells

Author: Goudy, Laine, Ha, Alvin, Borah, Ashir A., Umhoefer, Jennifer M., Chow, Lauren, Tran, Carinna, Winters, Aidan, Talbot, Alexis, Hernandez, Rosmely, Li, Zhongmei, Subramanya, Sanjana, Arab, Abolfazl, Kale, Nupura, Lee, Jae Hyun J., Muldoon, Joseph J., Liu, Chang, Schmidt, Ralf, Santangelo, Philip, Carnevale, Julia, Eyquem, Justin, Shy, Brian R., Marson, Alex, Gilbert, Luke A.

Issue&Volume: 2025-10-21

Abstract: Targeted epigenetic engineering of gene expression in cell therapies would allow programming of desirable phenotypes without many of the challenges and safety risks associated with double-strand break-based genetic editing approaches. Here, we develop an all-RNA platform for efficient, durable and multiplexed epigenetic programming in primary human T cells, stably turning endogenous genes off or on using CRISPRoff and CRISPRon epigenetic editors. We achieve epigenetic programming of diverse targeted genomic elements without the need for sustained expression of CRISPR systems. CRISPRoff-mediated gene silencing is maintained through numerous cell divisions, T cell stimulations and in vivo adoptive transfer, avoiding cytotoxicity or chromosomal abnormalities inherent to multiplexed Cas9-mediated genome editing. Lastly, we successfully combined genetic and epigenetic engineering using orthogonal CRISPR Cas12a–dCas9 systems for targeted chimeric antigen receptor (CAR) knock-in and CRISPRoff silencing of therapeutically relevant genes to improve preclinical CAR-T cell-mediated in vivo tumor control and survival.

DOI: 10.1038/s41587-025-02856-w

Source: https://www.nature.com/articles/s41587-025-02856-w