GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE338nnn/GSE338028/primaryOK200TranscriptomicsHomo sapiensExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE338028GEOGSEfalseLipid-based mRNA chimeric antigen receptor T cells for enhanced in vitro antitumor activityChimeric antigen receptor (CAR) T cell therapy provides substantial clinical benefits in hematologic malignancies. However, cell engineering using conventional viral vectors is costly, labor-intensive, and prone to safety risks, while electroporation-induced cellular damage limits manufacturing scalability. This study reports a safe, high-efficiency, nonviral platform for generating mRNA-engineered CAR-T cells. Herein, eight ionizable lipids are synthesized and formulated into lipid nanoparticles. Systematic screening and structural optimization identified Lipid 6 as the lead candidate. Transfection efficiency is validated by delivering eGFP mRNA into DC2.4, peripheral blood mononuclear, dendritic, and natural killer cells. Murine models confirm robust in vivo expression and a favorable safety profile. Lipid 6–mediated delivery of MUC1 mRNA generates MUC1–CAR-T cells characterized by high surface receptor expression and potent cytotoxicity against MUC1-positive tumor cells. These findings establish Lipid 6 as a feasible strategy for primary human T-cell engineering, facilitating functional CAR expression with minimal toxicity.2026/08/12GSE338028GSM9865020GSM9865019GSM9865018GSM9865017GSM9865022GSM986502124676338028Homo sapiens