{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T12:11:58Z","timestamp":1792239118913,"version":"4.4.0"},"reference-count":48,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T00:00:00Z","timestamp":1756425600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100017610","name":"Shenzhen Science and Technology Innovation Program","doi-asserted-by":"publisher","award":["KJZD20240903102703005"],"award-info":[{"award-number":["KJZD20240903102703005"]}],"id":[{"id":"10.13039\/501100017610","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82402147"],"award-info":[{"award-number":["82402147"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["advanced.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Adv Healthcare Materials"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Over the past decade, CAR\u2010T cell therapy has achieved remarkable success in treating hematological malignancies. However, traditional CAR\u2010T cell engineering employs viral vectors, which has several limitations. Additionally, the immunosuppressive tumor microenvironment, particularly mediated by the PD\u20101\/PD\u2010L1 pathway, significantly restricts CAR\u2010T cell efficacy. CRISPR\/Cas9\u2010mediated PD\u20101 knockout can enhance CAR\u2010T cell anti\u2010tumor activity, but traditional electroporation (EP) method often damages T cells. Herein, a novel lipid nanoparticles (LNPs)\u2010mediated delivery technology are introduced to engineer CAR\u2010T cells. The LNPs platform enables the simultaneous expression of CAR cassette and CRISPR\/Cas9 gene editor in T cells via co\u2010delivery of multiplex RNAs (CD19 CAR mRNA+Cas9 mRNA+sgRNA targeting PD\u20101). Importantly, LNPs exhibit higher transfection efficiency and superior cell viability compared to traditional electroporation method. The engineered CAR\u2010T cells with PD\u20101 knockout, which express anti\u2010CD19 CAR, can specifically kill CD19+ Nalm\u20106 tumor cells in vitro and display enhanced anti\u2010tumor activity in vivo. Furthermore, LNPs\u2010mediated co\u2010delivery of Cas9 mRNA and sgRNAs targeting PD\u20101, TRAC, and B2M enables triple\u2010knockout of T cells with high editing efficiencies (76% for PD\u20101, 86% for TRAC, and 80% for B2M), highlighting the ability for multiplex gene editing. This LNP\u2010mediated delivery strategy has great potentials for the development of safer and more efficacious CAR\u2010T cells.<\/jats:p>","DOI":"10.1002\/adhm.202501475","type":"journal-article","created":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T11:21:04Z","timestamp":1756466464000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Co\u2010Delivery of Multiple RNAs via Lipid Nanoparticles Enables Precise Gene Editing of CAR\u2010T Cells"],"prefix":"10.1002","volume":"15","author":[{"given":"Mengge","family":"Wang","sequence":"first","affiliation":[{"name":"Institute of Advanced Biotechnology, Institute of Homeostatic Medicine, and School of Medicine Southern University of Science and Technology  Shenzhen 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qibin","family":"Liao","sequence":"additional","affiliation":[{"name":"Shenzhen Third People's Hospital The Second Affiliated Hospital, Southern University of Science and Technology  Shenzhen 518112 China"},{"name":"State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University Guangzhou Medical University  Guangzhou 511436 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shimeng","family":"Bai","sequence":"additional","affiliation":[{"name":"Shenzhen Third People's Hospital The Second Affiliated Hospital, Southern University of Science and Technology  Shenzhen 518112 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyi","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute of Advanced Biotechnology, Institute of Homeostatic Medicine, and School of Medicine Southern University of Science and Technology  Shenzhen 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanzheng","family":"Peng","sequence":"additional","affiliation":[{"name":"Kidney Transplant Department (Liver Transplant Department), Transplant Center, National Clinical Research Center for Infectious Disease, Shenzhen Third People's Hospital, The Second Affiliated Hospital Southern University of Science and Technology  Shenzhen 518112 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengpeng","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute of Advanced Biotechnology, Institute of Homeostatic Medicine, and School of Medicine Southern University of Science and Technology  Shenzhen 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongzhou","family":"Lu","sequence":"additional","affiliation":[{"name":"National Clinical Research Center for Infectious Diseases, Shenzhen Third People's Hospital, The Second Affiliated Hospital Southern University of Science and Technology  Shenzhen 518112 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian\u2010Kang","family":"Zhu","sequence":"additional","affiliation":[{"name":"Institute of Advanced Biotechnology, Institute of Homeostatic Medicine, and School of Medicine Southern University of Science and Technology  Shenzhen 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5350-7639","authenticated-orcid":false,"given":"Chen","family":"Zeng","sequence":"additional","affiliation":[{"name":"Institute of Advanced Biotechnology, Institute of Homeostatic Medicine, and School of Medicine Southern University of Science and Technology  Shenzhen 518055 China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,8,29]]},"reference":[{"key":"e_1_2_9_1_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar6711"},{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra1706169"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41551-018-0235-9"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa2024850"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(21)00933-8"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-08333-8"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10529-010-0387-5"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1203162"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/iub.1566"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2017.098"},{"key":"e_1_2_9_11_1","author":"Dulery R.","year":"2025","journal-title":"Nat. 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