Automated Good Manufacturing Practice-Compatible CRISPR-Cas9 Editing of Hematopoietic Stem and Progenitor Cells for Clinical Treatment of β-Hemoglobinopathies
| العنوان: | Automated Good Manufacturing Practice-Compatible CRISPR-Cas9 Editing of Hematopoietic Stem and Progenitor Cells for Clinical Treatment of β-Hemoglobinopathies |
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| المؤلفون: | Guillermo Ureña-Bailén, Milena Block, Tommaso Grandi, Faidra Aivazidou, Jona Quednau, Dariusz Krenz, Alberto Daniel-Moreno, Andrés Lamsfus-Calle, Thomas Epting, Rupert Handgretinger, Stefan Wild, Markus Mezger |
| المصدر: | CRISPR J |
| بيانات النشر: | Mary Ann Liebert Inc, 2023. |
| سنة النشر: | 2023 |
| مصطلحات موضوعية: | Genetics, Original Research, Biotechnology |
| الوصف: | Cellular therapies hold enormous potential for the cure of severe hematological and oncological disorders. The forefront of innovative gene therapy approaches including therapeutic gene editing and hematopoietic stem cell transplantation needs to be processed by good manufacturing practice to ensure safe application in patients. In the present study, an effective transfection protocol for automated clinical-scale production of genetically modified hematopoietic stem and progenitor cells (HSPCs) using the CliniMACS Prodigy(®) system including the CliniMACS Electroporator (Miltenyi Biotec) was established. As a proof-of-concept, the enhancer of the BCL11A gene, clustered regularly interspaced short palindromic repeat (CRISPR) target in ongoing clinical trials for β-thalassemia and sickle-cell disease treatment, was disrupted by the CRISPR-Cas9 system simulating a large-scale clinical scenario, yielding 100 million HSPCs with high editing efficiency. In vitro erythroid differentiation and high-performance liquid chromatography analyses corroborated fetal hemoglobin resurgence in edited samples, supporting the feasibility of running the complete process of HSPC gene editing in an automated closed system. |
| تدمد: | 2573-1602 2573-1599 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bafb96aa4563ed81a540cfde5a2ab2e7 https://doi.org/10.1089/crispr.2022.0086 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....bafb96aa4563ed81a540cfde5a2ab2e7 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 25731602 25731599 |
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