This paper describes a precise method of gene titration as applied to the alpha- and beta-globin genes in the mouse. The three salient features of the method are: (i) the use of saturation hybridization in probe cDNA excess, (ii) the use of highly purified cDNA probes prepared by preparative hybridization with cloned globin sequences (Longacre and Mach (1978) J. Biol. Chem. 253, 7500) and (iii)...

We have analyzed the chromatin structure of the human beta-globin locus in somatic cell hybrids resulting from the fusion of human non-erythroid cells and mouse erythroleukemia (MEL) cells. In these hybrids, the human adult beta-globin gene, but neither the embryonic nor fetal globin genes, is activated transcriptionally. In addition, the DNase I-resistant beta-like globin locus characteristic ...

Hemoglobinopathies such as sickle cell anemia and b-thalassemia result from among the most common single gene defects worldwide. A promising approach for the treatment of these conditions is through the induction of increased fetal hemoglobin (HbF) expression. Hydroxyurea, which is currently part of the standard treatment of sickle cell anemia, causes increased expression of HbF. However, the l...

Functional human globin messenger RNA but failed to stimulate the synthesis of any was isolated from reticulocytes of two paf. -chains, even though nonthalassemic tients with homozygous /3#{176}-thalassemia, human globin mRNA preparations conthree patients with sickle cell fl#{176}-thalassistently stimulated two to four times as semia, and one patient doubly heteromuch fl or globin chain synthe...

We sought to investigate the usefulness of the adeno-associated virus 2 (AAV)-based vectors to suppress the excess production of the human alpha-globin gene product towards developing a treatment modality for beta-thalassemia since accumulation of free alpha-globin reduces the lifespan of red blood cells in these patients. We constructed recombinant AAV virions containing the human alpha-globin...

The development and evaluation of drugs to elevate fetal hemoglobin in the treatment of the genetic diseases of hemoglobin would be facilitated by the availability of reliable cell assays. We have used real-time, quantitative polymerase chain reaction (PCR) analyses of globin messenger RNA (mRNA) levels in a biphasic, erythropoietin-dependent primary culture system for human adult erythroid cel...

Beta thalassemia is the  most common autosomal recessive disorder. The present study reports a rare β globin gene mutation, HBB: c.180G>A: codon 59 (AAG/AAA), in a patient from Gilan province, northern Iran. Nucleotide sequencing of amplified DNA belonging to a 35 years old man presenting mild hypochromia revealed a synonymous mutation due to a G>A conversion at the third position of codon 59 o...

In order to test whether the ontogenetic origin of human parental cells influences the expression of globin genes, we did fusions of mouse erythroleukemia (MEL) cells with fetal or adult nonerythroid cells and we assessed globin expression in hybrids. For selection of hybrid clones we used a monoclonal antibody that recognizes a human chromosome 11-linked cell surface marker. Globin expression ...

In order to test if trans-acting regulatory factors specific for globin genes of the adult and embryonic stages of development exist in erythroid cells, transcriptionally active embryonic and adult globin genes on the same chromosome were transferred by cell fusion from the human leukemia cell K562 into phenotypically adult mouse erythroleukemia cells. Restriction-fragment-length polymorphisms ...