Year: 2025
Type: article
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.
Source: Nature Biotechnology
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
Institutions: Gladstone Institutes, University of California, San Francisco, Arc Research Institute, Inserm, Université Paris Cité, Hôpital Saint-Louis, Medical University of Vienna, Georgia Institute of Technology, The Wallace H. Coulter Department of Biomedical Engineering, Parker Institute for Cancer Immunotherapy, UCSF Helen Diller Family Comprehensive Cancer Center, Innovative Genomics Institute, University of California, Berkeley
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $12,690
Funders: National Science Foundation, U.S. Department of Health and Human Services, Cancer Research Institute, Parker Institute for Cancer Immunotherapy, Emerson Collective, National Institutes of Health, University of California, San Francisco, National Human Genome Research Institute, National Cancer Institute
Awards: G1586206470, Non-viral genome, epigenome, and transcriptome engineering for clinical CAR-T cell manufacturing, G4099230581, G4482867767, G4680438090, G4785822992, Targeting SOCS1 and RASA2 to Engineer More Potent Adoptive T Cell Therapies for Cancer Treatment., G5463808913, G7306057323, G9000032459, Spatial multiomic mapping of gene function with CRISPRoff
OpenAlex ID: W4415385405