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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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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. |
format | Online Article Text |
id | pubmed-9601852 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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