ORIGINAL ARTICLES  TRENDS IN MOLECULAR DIAGNOSIS AND THERAPY OF β-THALASSEMIA AND SICKLE CELL ANEMIA 

Minerva Biotecnologica 2003 June;15(2):137-44

Copyright © 2003 EDIZIONI MINERVA MEDICA

language: English

Quantitative RT-PCR for the analysis of expression of α-, β- and γ-globin genes in erythroid cells

Bianchi N. ^{1, 2}, Borgatti N. ^{1, 2}, Gambari R. ^{1, 2}

¹ Department of Biochemistry and Molecular Biology, University of Ferrara, Ferrara, Italy; ² Laboratory for the Development of Pharmacological and Pharmacogenomic Therapy of Thalassemia, Biotechnology Center, University of Ferrara, Ferrara, Italy

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The identification of molecules useful for pharmacologically-mediated regulation of the expression of human γ-globin genes is a crucial step for the development of agents potentially useful in experimental therapy of hematological disorders, including β-thalassemia and sickle cell anemia. It is well established, indeed, that increase of fetal hemoglobin (HbF) to 30% of the total hemoglobin (Hb) leads to a significant improvement of the clinical status of patients affected by these hematological disorders. In this respect, a rapid and reproducible analysis of the effects of potential HbF inducers on accumulation of mRNAs for γ-, β- and α-globins is relevant. Here, we review data on the use of real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) for studying accumulation of mRNAs for γ-, β- and α-globins. By this procedure it is possible to differentiate inducers able to stimulate preferential expression of γ-globin genes from inducers able to stimulate both γ-globin and β-globin mRNA production. This analytical strategy is of potential clinical significance to identify HbF inducers useful to alleviate the symptoms underlying β-thalassemia and sickle cell anemia, and compounds stimulating β-globin gene expression, of interest in a variety of β+-thalassemias.