Beta-Globin Gene Cluster Haplotypes in Yemeni Children with Sickle Cell Disease

2010 ◽  
Vol 123 (3) ◽  
pp. 182-185 ◽  
Author(s):  
Abdul-Wahab M. Al-Saqladi ◽  
Bernard J. Brabin ◽  
Hassan A. Bin-Gadeem ◽  
Warsha A. Kanhai ◽  
Marion Phylipsen ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Gene Cluster ◽  
Sickle Cell ◽  
Globin Gene ◽  
Beta Globin ◽  
Cell Disease ◽  
Beta Globin Gene ◽  
Globin Gene Cluster

Rapid and reliable β-globin gene cluster haplotyping of sickle cell disease patients by FRET Light Cycler and HRM assays

2011 ◽  
Vol 412 (13-14) ◽  
pp. 1257-1261 ◽  
Author(s):  
Philippe Joly ◽  
Philippe Lacan ◽  
Caroline Garcia ◽  
Angelique Delasaux ◽  
Alain Francina
Keyword(s):  
Sickle Cell Disease ◽  
Gene Cluster ◽  
Sickle Cell ◽  
Globin Gene ◽  
Cell Disease ◽  
Globin Gene Cluster ◽  
Light Cycler

scholarly journals Production of F cells in sickle cell anemia: regulation by a genetic locus or loci separate from the beta-globin gene cluster

Blood ◽  
1984 ◽  
Vol 64 (5) ◽  
pp. 1053-1058 ◽  
Author(s):  
SH Boyer ◽  
GJ Dover ◽  
GR Serjeant ◽  
KD Smith ◽  
SE Antonarakis ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Sickle Cell ◽  
Genetic Locus ◽  
Globin Gene ◽  
The United States ◽  
Beta Globin ◽  
Cell Production ◽  
Beta Globin Gene ◽  
Globin Gene Cluster ◽  
Sib Pairs

Abstract Levels of fetal hemoglobin (HbF) bearing reticulocytes (F reticulocytes) range from 2% to 50% in patients with sickle cell (SS) anemia. To learn whether any portion of such variation in F cell production is regulated by loci genetically separable from the beta- globin gene cluster, percentages of F reticulocytes were compared in 59 sib pairs composed solely of SS members, including 40 pairs from Jamaica and 19 from the United States. We reasoned that differences in F reticulocyte levels might arise (1) from any of several kinds of artifact, (2) via half-sib status, or (3) because one or more genes regulating F cell production segregate separately from beta S. We minimized the role of artifact by assay of fresh samples from 84 SS individuals, including both members of 38 sib pairs. In 78 of the 84 subjects, serial values for percent F reticulocytes fell within 99.9% confidence limits or were alike by t test (P greater than or equal to .05). This left 32 sib pairs for which F reticulocyte levels in each member were reproducible. When sib-sib comparisons were limited to these 32 pairs, percentages of F reticulocytes were grossly dissimilar within 12 Jamaican and 3 American sibships. Within them, the probability that sibs were alike was always less than or equal to .005 and usually less than or equal to 10(-4). We next minimized the contribution of half-sibs among Jamaicans by a combination of paternity testing and sib-sib comparison of beta-globin region DNA restriction fragment length polymorphisms, especially among discordant pairs. We thereafter concluded that at least seven to eight Jamaican pairs were composed of reproducibly discordant full sibs. There is thus little doubt that there are genes regulating between-patient differences in F cell production that are separate from the beta-globin gene cluster. Still unanswered is (1) whether or not these genes are actually linked to beta S, (2) why F reticulocyte levels in Americans tend to be lower than in Jamaicans, and (3) whether or not differences in F cell production among SS patients are regulated by several major loci or by only one.

β-Globin gene cluster haplotypes and HbF levels are not the only modulators of sickle cell disease in Lebanon

2003 ◽  
Vol 70 (2) ◽  
pp. 79-83 ◽  
Author(s):  
A. Inati ◽  
A. Taher ◽  
W. Bou Alawi W ◽  
S. Koussa ◽  
H. Kaspar ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Gene Cluster ◽  
Sickle Cell ◽  
Globin Gene ◽  
Cell Disease ◽  
Globin Gene Cluster

scholarly journals Haplotypes of the beta-globin gene as prognostic factors in sickle-cell disease

1999 ◽  
Vol 5 (6) ◽  
pp. 1254-1258
Author(s):  
M. A. El Hazmi ◽  
A. S. Warsy ◽  
N. Bashir ◽  
A. Beshlawi ◽  
I. R. Hussain
Keyword(s):  
Saudi Arabia ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Clinical Presentation ◽  
Globin Gene ◽  
Beta Globin ◽  
Cell Disease ◽  
Beta Globin Gene ◽  
The North ◽  
Cell Gene

Wecollaborated with researchers from Egypt, Syrian Arab Republic and Jordan in a study of patients with sickle-cell disease from those countries, and from various parts of Saudi Arabia, in order to investigate the influence of genetics on the clinical presentation of the disease, and to attempt to determine the origin of the sickle-cell gene in Arabs. Our results suggest that beta-globin gene haplotypes influence the clinical presentation of sickle-cell disease, and that there are at least two major foci for the origin of the sickle-cell gene, one in the eastern part of Saudi Arabia, and the other in the populations of North Africa and the north-western part of the Arabian peninsula

Target Site Mutagenesis during Crispr/ Cas 9/Single-Stranded- Oligonucleotide Directed Gene Editing for Sickle Cell Anemia

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 4706-4706 ◽  
Author(s):  
Dula Man ◽  
Brett Sansbury ◽  
Pawel Bialk ◽  
Kevin Bloh ◽  
E. Anders Kolb ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Sanger Sequencing ◽  
Gene Editing ◽  
Globin Gene ◽  
Beta Globin ◽  
Cell Disease ◽  
Beta Globin Gene ◽  
Site Mutagenesis ◽  
Single Stranded Oligonucleotide

Abstract Introduction Sickle Cell Disease (SCD) results from a simple substitution of valine for glutamic acid at codon 6 in the β globin gene, resulting in an AàT transition in the third position. The mutation results in the production of hemoglobin HbS which differs from the normal HbAin that it tends to polymerize into long strands that deform the erythrocyte. While a variety of traditional treatment regimens or reagents, such as hydroxyurea and chronic transfusions have been used widely, these therapies are wrought with short and long term side effects that limit efficacy. There is great interest in the hypothesis that the repair of a single nucleotide is facilitated by the combined action of CRISPR/Cas 9 and single-stranded oligonucleotides (ssODNs) could prove a significant therapeutic advance for sickle cell disease. CRISPR/Cas 9 induces a site-specific double-stranded break while the single-stranded oligonucleotide provides a DNA template to improve the rate of accurate genetic correction. There is a great effort to understand off-site mutagenesis caused by editing of DNA regions remote to the target sequence. It is critical that investigators understand the frequency and types of DNA alterations induced by the gene editing reaction and affecting the region surrounding the targeted nucleotide site (herein termed on-site mutagenesis). Methods We targeted beta globin genes with a CRISPR/Cas9 system designed to cleave 2 bases to the 5'side of the targeted (A) nucleotide (the PAM site is located 2 bases to the 3' side on the complimentary strand) coupled to the electroporation of a single-stranded oligonucleotide (72-mer) designed to convert the wild-type (A) to the mutant (T) nucleotide. To evaluate the rate of on-site mutagenesis among individual alleles, we clonally expanded populations of edited K562 cells. We hypothesize that the evaluation of individual clones will permit a clear identification of intended and un-intended on-site DNA alteration. Allelic heterogeneity is analyzed using Tracking of Indels by DEcompositon (TIDE) methodology combined with Sanger sequencing. Results We isolated 26 clonal lines for continued expansion after evidence of CRISPR/Cas 9 plasmid uptake and activity. Sanger sequencing with TIDE analysis revealed that the DNA sequence surrounding the targeted base is altered significantly as a result of the CRISPR/Cas9 gene editing process. Twenty three percent of the clones contain at least one corrected allele but one hundred percent of the clones exhibited mutagenicity of the DNA sequence surrounding the targeted base. All clones analyzed displayed varying degrees of sequence alteration (Figure 1). Interestingly, one clone contained a DNA insertion homologous to a region of the delta globin gene, suggesting that gene editing of the beta globin gene may have been repaired, in part, by delta globin DNA. The sequence of the delta globin locus was not changed. Conclusions Taken together, our data suggest that combinatorial approaches to beta globin gene editing using CRISPR/Cas 9 and single-stranded oligonucleotides induce significant onsite mutagenesis and potential genetic swapping between related members of the same gene family. These observations provide insight into the type of molecular activity that accompanies combinatorial gene editing, particularly surrounding the target site. This work is supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number P20GM109021 Disclosures No relevant conflicts of interest to declare.

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