Integrated epigenetic and genetic programming of primary human T cells.

IF 41.7 1区生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Nature biotechnology Pub Date : 2025-10-21 DOI:10.1038/s41587-025-02856-w

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

{"title":"Integrated epigenetic and genetic programming of primary human T cells.","authors":"Laine Goudy,Alvin Ha,Ashir A Borah,Jennifer M Umhoefer,Lauren Chow,Carinna Tran,Aidan Winters,Alexis Talbot,Rosmely Hernandez,Zhongmei Li,Sanjana Subramanya,Abolfazl Arab,Nupura Kale,Jae Hyun J Lee,Joseph J Muldoon,Chang Liu,Ralf Schmidt,Philip Santangelo,Julia Carnevale,Justin Eyquem,Brian R Shy,Alex Marson,Luke A Gilbert","doi":"10.1038/s41587-025-02856-w","DOIUrl":null,"url":null,"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.","PeriodicalId":19084,"journal":{"name":"Nature biotechnology","volume":"1 1","pages":""},"PeriodicalIF":41.7000,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nature biotechnology","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1038/s41587-025-02856-w","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}

引用次数: 0

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.

查看原文本刊更多论文

人类原代T细胞的综合表观遗传和遗传编程。

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。在这里，我们开发了一个全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，使用CRISPRoff和CRISPRon表观遗传编辑器稳定地关闭或打开内源性基因。我们实现了多种靶向基因组元件的表观遗传编程，而不需要持续表达CRISPR系统。crisprof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，我们成功地结合了基因和表观遗传工程，使用正交CRISPR Cas12a-dCas9系统进行靶向嵌合抗原受体（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。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Nature biotechnology 工程技术-生物工程与应用微生物

CiteScore

63.00

自引率

1.70%

发文量

382

审稿时长

3 months

期刊介绍： Nature Biotechnology is a monthly journal that focuses on the science and business of biotechnology. It covers a wide range of topics including technology/methodology advancements in the biological, biomedical, agricultural, and environmental sciences. The journal also explores the commercial, political, ethical, legal, and societal aspects of this research. The journal serves researchers by providing peer-reviewed research papers in the field of biotechnology. It also serves the business community by delivering news about research developments. This approach ensures that both the scientific and business communities are well-informed and able to stay up-to-date on the latest advancements and opportunities in the field. Some key areas of interest in which the journal actively seeks research papers include molecular engineering of nucleic acids and proteins, molecular therapy, large-scale biology, computational biology, regenerative medicine, imaging technology, analytical biotechnology, applied immunology, food and agricultural biotechnology, and environmental biotechnology. In summary, Nature Biotechnology is a comprehensive journal that covers both the scientific and business aspects of biotechnology. It strives to provide researchers with valuable research papers and news while also delivering important scientific advancements to the business community.