CRISPR-mediated gene modification of hematopoietic stem cells with beta-thalassemia IVS-1-110 mutation
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RESEARCH
Open Access
CRISPR-mediated gene modification of hematopoietic stem cells with betathalassemia IVS-1-110 mutation Hala Gabr1, Mona Kamal El Ghamrawy2, Abdulrahman H. Almaeen3, Ahmed Samir Abdelhafiz4, Aya Osama Saad Hassan1 and Maha Hamdi El Sissy1*
Abstract Background: β-Thalassemias represent a group of genetic disorders caused by human hemoglobin beta (HBB) gene mutations. The radical curative approach is to correct the mutations causing the disease. CRISPR-CAS9 is a novel gene-editing technology that can be used auspiciously for the treatment of these disorders. The study aimed to investigate the utility of CRISPR-CAS9 for gene modification of hematopoietic stem cells in β-thalassemia with IVS-1-110 mutation. Methods and results: We successfully isolated CD34+ cells from peripheral blood of β-thalassemia patients with IVS-1-110 mutation. The cells were transfected with Cas9 endonuclease together with guide RNA to create doublestrand breaks and knock out the mutation. The mutation-corrected CD34+ cells were subjected to erythroid differentiation by culturing in complete media containing erythropoietin. Conclusion: CRISPR/Cas-9 is an effective tool for gene therapy that will broaden the spectrum of therapy and potentially improve the outcomes of β-thalassemia. Keywords: Thalassemia, CRISPR/Cas-9, Reverse hybridization, Hemoglobin beta gene mutation, Hematopoietic stem cells
Background Mutations involving the β-globin gene are the most common cause of genetic disorders in humans. βThalassemia constitutes a group of hereditary disorders affecting the expression of adult β-globin gene. They are caused by a few hundred of mutations decreasing or completely negating β-globin production. As a result, adult α2β2 hemoglobin (HbA) is reduced and excess α-globin content accumulates in the erythroid cells resulting in ineffective erythropoiesis and augmented apoptosis [20]. Besides being prevalent * Correspondence: [email protected] 1 Clinical and Chemical Pathology, Faculty of Medicine, Cairo University, Cairo, Egypt Full list of author information is available at the end of the article
among Africans, β-thalassemias are also common in Greeks and Italians [30]. It has been estimated that 1000 children out of 1.5 million live births are born annually with thalassemia major in Egypt [11]. In Egyptian multi-centric studies, the carrier rate has been reported to be in the range of 9–10% [9]. The more than 350 β-thalassemia mutations reported are geographically widely diverse with a small number of specific mutations in individual populations. Thalassemia syndromes are common in Saudi Arabia. Betathalassemia gene mutations, particularly B+ and Bo thalassemia, occur with variable frequency in different regions of Saudi Arabia [6]. Thalassemia therapy constitutes a major socioeconomic burden. Various methods of treatment have been
© The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and rep
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