TY  - JOUR
JF  - Molecular Therapy: Methods & Clinical Development
A1  - Woodruff, Rosie
A1  - Parekh, Farhaan
A1  - Lamb, Katarina
A1  - Mekkaoui, Leila
A1  - Allen, Christopher
A1  - Smetanova, Katerina
A1  - Huang, Jasmine
A1  - Williams, Alex
A1  - Toledo, Gerardo Santiago
A1  - Lilova, Koki
A1  - Roddie, Claire
A1  - Sillibourne, James
A1  - Pule, Martin
KW  - base editing
KW  -  BE4max
KW  -  chimeric antigen receptor
KW  -  circular RNAPD-1TIM3
N2  - 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 × 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 ?g circRNA/1 × 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.
ID  - discovery10180184
PB  - Elsevier BV
UR  - https://doi.org/10.1016/j.omtm.2023.101123
N1  - © 2023 The Authors. Published by Elsevier under a Creative Commons license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
TI  - Large-scale manufacturing of base-edited chimeric antigen receptor T cells
AV  - public
VL  - 31
Y1  - 2023/12/14/
ER  -