@article{discovery10180184,
          volume = {31},
            note = {{\copyright} 2023 The Authors. Published by Elsevier under a Creative Commons license (http://creativecommons.org/licenses/by-nc-nd/4.0/).},
           title = {Large-scale manufacturing of base-edited chimeric antigen receptor T�cells},
           month = {December},
         journal = {Molecular Therapy: Methods \& Clinical Development},
            year = {2023},
       publisher = {Elsevier BV},
             url = {https://doi.org/10.1016/j.omtm.2023.101123},
        keywords = {base editing, BE4max, chimeric antigen receptor, circular RNAPD-1TIM3},
        abstract = {Base editing is a revolutionary gene-editing technique enabling the introduction of point mutations into the genome without generating detrimental DNA double-stranded breaks. Base-editing enzymes are commonly delivered in the form of modified linear messenger RNA (mRNA) that is costly to produce. Here, we address this problem by developing a simple protocol for manufacturing base-edited cells using circular RNA (circRNA), which is less expensive to synthesize. Compared with linear mRNA, higher editing efficiencies were achieved with circRNA, enabling an 8-fold reduction in the amount of RNA required. We used this protocol to manufacture a clinical dose (1�{$\times$}�108 cells) of base-edited chimeric antigen receptor (CAR) T�cells lacking expression of the inhibitory receptor, PD-1. Editing efficiencies of up to 86\% were obtained using 0.25�{\ensuremath{\mu}}g circRNA/1�{$\times$}�106 cells. Increased editing efficiencies with circRNA were attributed to more efficient translation. These results suggest that circRNA, which is less expensive to produce than linear mRNA, is a viable option for reducing the cost of manufacturing base-edited cells at scale.},
          author = {Woodruff, Rosie and Parekh, Farhaan and Lamb, Katarina and Mekkaoui, Leila and Allen, Christopher and Smetanova, Katerina and Huang, Jasmine and Williams, Alex and Toledo, Gerardo Santiago and Lilova, Koki and Roddie, Claire and Sillibourne, James and Pule, Martin}
}