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Large-scale manufacturing of base-edited chimeric antigen receptor T cells
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. He...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10597784/ https://www.ncbi.nlm.nih.gov/pubmed/37886606 http://dx.doi.org/10.1016/j.omtm.2023.101123 |
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author | Woodruff, Rosie Parekh, Farhaan Lamb, Katarina Mekkaoui, Leila Allen, Christopher Smetanova, Katerina Huang, Jasmine Williams, Alex Toledo, Gerardo Santiago Lilova, Koki Roddie, Claire Sillibourne, James Pule, Martin |
author_facet | Woodruff, Rosie Parekh, Farhaan Lamb, Katarina Mekkaoui, Leila Allen, Christopher Smetanova, Katerina Huang, Jasmine Williams, Alex Toledo, Gerardo Santiago Lilova, Koki Roddie, Claire Sillibourne, James Pule, Martin |
author_sort | Woodruff, Rosie |
collection | PubMed |
description | 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 × 10(8) 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 μg circRNA/1 × 10(6) 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. |
format | Online Article Text |
id | pubmed-10597784 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-105977842023-10-26 Large-scale manufacturing of base-edited chimeric antigen receptor T cells Woodruff, Rosie Parekh, Farhaan Lamb, Katarina Mekkaoui, Leila Allen, Christopher Smetanova, Katerina Huang, Jasmine Williams, Alex Toledo, Gerardo Santiago Lilova, Koki Roddie, Claire Sillibourne, James Pule, Martin Mol Ther Methods Clin Dev Original Article 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 × 10(8) 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 μg circRNA/1 × 10(6) 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. American Society of Gene & Cell Therapy 2023-10-05 /pmc/articles/PMC10597784/ /pubmed/37886606 http://dx.doi.org/10.1016/j.omtm.2023.101123 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Woodruff, Rosie Parekh, Farhaan Lamb, Katarina Mekkaoui, Leila Allen, Christopher Smetanova, Katerina Huang, Jasmine Williams, Alex Toledo, Gerardo Santiago Lilova, Koki Roddie, Claire Sillibourne, James Pule, Martin Large-scale manufacturing of base-edited chimeric antigen receptor T cells |
title | Large-scale manufacturing of base-edited chimeric antigen receptor T cells |
title_full | Large-scale manufacturing of base-edited chimeric antigen receptor T cells |
title_fullStr | Large-scale manufacturing of base-edited chimeric antigen receptor T cells |
title_full_unstemmed | Large-scale manufacturing of base-edited chimeric antigen receptor T cells |
title_short | Large-scale manufacturing of base-edited chimeric antigen receptor T cells |
title_sort | large-scale manufacturing of base-edited chimeric antigen receptor t cells |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10597784/ https://www.ncbi.nlm.nih.gov/pubmed/37886606 http://dx.doi.org/10.1016/j.omtm.2023.101123 |
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