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Iranian Journal of Basic Medical Sciences، جلد ۱۹، شماره ۱۰، صفحات ۱۰۶۳-۱۰۶۹
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عنوان انگلیسی Gamma reactivation using the spongy effect of KLF1-binding site sequence: an approach in gene therapy for beta-thalassemia
چکیده انگلیسی مقاله Objective(s): β-thalassemia is one of the most common genetic disorders in the world. As one of the promising treatment strategies, fetal hemoglobin (Hb F) can be induced. The present study was an attempt to reactivate the γ-globin gene by introducing a gene construct containing KLF1 binding sites to the K562 cell line. Materials and Methods: A plasmid containing a 192 bp sequence with two repeats of KLF1 binding sites on β-globin and BCL11A promoters was constructed and used to transfect the K562 cell line. Positive selection was performed under treatment with 150 μg/ml hygromycin B. The remaining cells were expanded and harvested on day 28, and genomic DNA was extracted. The PCR was carried out to verify insertion of DNA fragment to the genome of K562 cells. The cells were differentiated with 15 µg/ml cisplatin. Flowcytometry was performed to identify erythroid differentiation by detection of CD235a+ cells. Real-time RT-PCR was performed to evaluate γ-globin expression in the transfected cells. Results: A 1700 bp fragment was observed on agarose gel as expected and insertion of DNA fragment to the genome of K562 cells was verified. Totally, 84% of cells were differentiated. The transfected cells significantly increased γ-globin expression after differentiation compared to untransfected ones. Conclusion: The findings demonstrate that the spongy effect of KLF1-binding site on BCL11A and β-globin promoters can induce γ-globin expression in K562 cells. This novel strategy can be promising for the treatment of β-thalassemia and sickle cell disease.
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نویسندگان مقاله نسرین حیدری | nasrin heydari
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

لاله شریعتی | laleh shariati
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

حسین خان احمد | hossein khanahmad
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran|pediatric inherited diseases research center, research institute for primordial prevention of non-communicable disease, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

زهرا حجازی | zahra hejazi
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

منصوره شهبازی | mansoureh shahbazi
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

منصور صالحی | mansoor salehi
department of genetics and molecular biology, school of medicine, isfahan university of medical sciences, isfahan, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اصفهان (Isfahan university of medical sciences)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_7731.html
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زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Article
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