Isolation and translation of hemoglobin messenger RNA from thalassemia, sickle cell anemia, and normal human reticulocytes

Human hemoglobin messenger RNA was isolated by sucrose gradient centrifugation from reticulocytes of patients having various hemolytic anemias. Using a messenger RNA-dependent cell-free system derived entirely from rabbit reticulocytes, the human hemoglobin messenger RNA has been translated and the products analyzed by carboxymethylcellulose column chromatography. Normal messenger RNA directs synthesis of normal human alpha- and beta-globin chains in nearly equal amounts. Sickle cell anemia messenger RNA directs the synthesis of normal alpha- and sickle beta-chains, beta-thalassemia messenger RNA directs the synthesis of normal alpha- and beta-chains, but the amount of beta-globin synthesized is markedly reduced. Thus the inability of the thalassemia reticulocyte to produce beta-globin is clearly attributable to the beta-globin messenger RNA.