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Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin

Gene editing (GE) is an efficient strategy for correcting genetic mutations in monogenic hereditary diseases, including β-thalassemia. We have elsewhere reported that CRISPR-Cas9-based gene editing can be employed for the efficient correction of the β(0)39-thalassemia mutation. On the other hand, ro...

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Autores principales: Cosenza, Lucia Carmela, Zuccato, Cristina, Zurlo, Matteo, Gambari, Roberto, Finotti, Alessia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9601852/
https://www.ncbi.nlm.nih.gov/pubmed/36292612
http://dx.doi.org/10.3390/genes13101727
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author Cosenza, Lucia Carmela
Zuccato, Cristina
Zurlo, Matteo
Gambari, Roberto
Finotti, Alessia
author_facet Cosenza, Lucia Carmela
Zuccato, Cristina
Zurlo, Matteo
Gambari, Roberto
Finotti, Alessia
author_sort Cosenza, Lucia Carmela
collection PubMed
description Gene editing (GE) is an efficient strategy for correcting genetic mutations in monogenic hereditary diseases, including β-thalassemia. We have elsewhere reported that CRISPR-Cas9-based gene editing can be employed for the efficient correction of the β(0)39-thalassemia mutation. On the other hand, robust evidence demonstrates that the increased production of fetal hemoglobin (HbF) can be beneficial for patients with β-thalassemia. The aim of our study was to verify whether the de novo production of adult hemoglobin (HbA) using CRISPR-Cas9 gene editing can be combined with HbF induction protocols. The gene editing of the β(0)39-globin mutation was obtained using a CRISPR-Cas9-based experimental strategy; the correction of the gene sequence and the transcription of the corrected gene were analyzed by allele-specific droplet digital PCR and RT-qPCR, respectively; the relative content of HbA and HbF was studied by high-performance liquid chromatography (HPLC) and Western blotting. For HbF induction, the repurposed drug rapamycin was used. The data obtained conclusively demonstrate that the maximal production of HbA and HbF is obtained in GE-corrected, rapamycin-induced erythroid progenitors isolated from β(0)39-thalassemia patients. In conclusion, GE and HbF induction might be used in combination in order to achieve the de novo production of HbA together with an increase in induced HbF.
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spelling pubmed-96018522022-10-27 Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin Cosenza, Lucia Carmela Zuccato, Cristina Zurlo, Matteo Gambari, Roberto Finotti, Alessia Genes (Basel) Article Gene editing (GE) is an efficient strategy for correcting genetic mutations in monogenic hereditary diseases, including β-thalassemia. We have elsewhere reported that CRISPR-Cas9-based gene editing can be employed for the efficient correction of the β(0)39-thalassemia mutation. On the other hand, robust evidence demonstrates that the increased production of fetal hemoglobin (HbF) can be beneficial for patients with β-thalassemia. The aim of our study was to verify whether the de novo production of adult hemoglobin (HbA) using CRISPR-Cas9 gene editing can be combined with HbF induction protocols. The gene editing of the β(0)39-globin mutation was obtained using a CRISPR-Cas9-based experimental strategy; the correction of the gene sequence and the transcription of the corrected gene were analyzed by allele-specific droplet digital PCR and RT-qPCR, respectively; the relative content of HbA and HbF was studied by high-performance liquid chromatography (HPLC) and Western blotting. For HbF induction, the repurposed drug rapamycin was used. The data obtained conclusively demonstrate that the maximal production of HbA and HbF is obtained in GE-corrected, rapamycin-induced erythroid progenitors isolated from β(0)39-thalassemia patients. In conclusion, GE and HbF induction might be used in combination in order to achieve the de novo production of HbA together with an increase in induced HbF. MDPI 2022-09-26 /pmc/articles/PMC9601852/ /pubmed/36292612 http://dx.doi.org/10.3390/genes13101727 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cosenza, Lucia Carmela
Zuccato, Cristina
Zurlo, Matteo
Gambari, Roberto
Finotti, Alessia
Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin
title Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin
title_full Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin
title_fullStr Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin
title_full_unstemmed Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin
title_short Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β(0)39-Globin Gene Editing and Induction of Fetal Hemoglobin
title_sort co-treatment of erythroid cells from β-thalassemia patients with crispr-cas9-based β(0)39-globin gene editing and induction of fetal hemoglobin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9601852/
https://www.ncbi.nlm.nih.gov/pubmed/36292612
http://dx.doi.org/10.3390/genes13101727
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