Expression analysis of gamma globin gene switching associated micro RNAs, flowing BCL11A gene inhibition

Gene Reports ◽  
2020 ◽  
Vol 21 ◽  
pp. 100934
Author(s):  
Narges Obeidi ◽  
Gholamhossein Tamaddon ◽  
Reza Ranjbaran ◽  
Gholamreza Khamisipour ◽  
Fatemeh Saberi
Keyword(s):  
Expression Analysis ◽  
Globin Gene ◽  
Gamma Globin ◽  
Micro Rnas ◽  
Gene Switching ◽  
Gene Inhibition

scholarly journals Analysis of mice containing a targeted deletion of beta-globin locus control region 5' hypersensitive site 3.

1996 ◽  
Vol 16 (6) ◽  
pp. 2906-2912 ◽  
Author(s):  
B A Hug ◽  
R L Wesselschmidt ◽  
S Fiering ◽  
M A Bender ◽  
E Epner ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Germ Line ◽  
Globin Gene ◽  
Globin Genes ◽  
Beta Globin ◽  
Erythroid Cells ◽  
Hypersensitive Site ◽  
Targeted Deletion ◽  
Gamma Globin ◽  
Gene Switching

To examine the function of murine beta-globin locus region (LCR) 5' hypersensitive site 3 (HS3) in its native chromosomal context, we deleted this site from the mouse germ line by using homologous recombination techniques. Previous experiments with human 5' HS3 in transgenic models suggested that this site independently contains at least 50% of total LCR activity and that it interacts preferentially with the human gamma-globin genes in embryonic erythroid cells. However, in this study, we demonstrate that deletion of murine 5' HS3 reduces expression of the linked embryonic epsilon y- and beta H 1-globin genes only minimally in yolk sac-derived erythroid cells and reduces output of the linked adult beta (beta major plus beta minor) globin genes by approximately 30% in adult erythrocytes. When the selectable marker PGK-neo cassette was left within the HS3 region of the LCR, a much more severe phenotype was observed at all developmental stages, suggesting that PGK-neo interferes with LCR activity when it is retained within the LCR. Collectively, these results suggest that murine 5' HS3 is not required for globin gene switching; importantly, however, it is required for approximately 30% of the total LCR activity associated with adult beta-globin gene expression in adult erythrocytes.

scholarly journals Human gamma- to beta-globin gene switching using a mini construct in transgenic mice.

1992 ◽  
Vol 12 (4) ◽  
pp. 1561-1567 ◽  
Author(s):  
J A Lloyd ◽  
J M Krakowsky ◽  
S C Crable ◽  
J B Lingrel
Keyword(s):  
Transgenic Mice ◽  
Developmental Regulation ◽  
Globin Gene ◽  
Globin Genes ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Gamma Globin ◽  
High Level ◽  
Gene Switching ◽  
Globin Gene Expression

The developmental regulation of the human globin genes involves a key switch from fetal (gamma-) to adult (beta-) globin gene expression. It is possible to study the mechanism of this switch by expressing the human globin genes in transgenic mice. Previous work has shown that high-level expression of the human globin genes in transgenic mice requires the presence of the locus control region (LCR) upstream of the genes in the beta-globin locus. High-level, correct developmental regulation of beta-globin gene expression in transgenic mice has previously been accomplished only in 30- to 40-kb genomic constructs containing the LCR and multiple genes from the locus. This suggests that either competition for LCR sequences by other globin genes or the presence of intergenic sequences from the beta-globin locus is required to silence the beta-globin gene in embryonic life. The results presented here clearly show that the presence of the gamma-globin gene (3.3 kb) alone is sufficient to down-regulate the beta-globin gene in embryonic transgenic mice made with an LCR-gamma-beta-globin mini construct. The results also show that the gamma-globin gene is down-regulated in adult mice from most transgenic lines made with LCR-gamma-globin constructs not including the beta-globin gene, i.e., that the gamma-globin gene can be autonomously regulated. Evidence presented here suggests that a region 3' of the gamma-globin gene may be important for down-regulation in the adult. The 5'HS2 gamma en beta construct described is a suitable model for further study of the mechanism of human gamma- to beta-globin gene switching in transgenic mice.

KLF1 Dependent Pathways In Developmental Globin Gene Switching

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 5162-5162
Author(s):  
Alexander E. Felice
Keyword(s):  
Conflicts Of Interest ◽  
Expression Profiles ◽  
Globin Gene ◽  
Fetal Hemoglobin ◽  
Erythroid Progenitors ◽  
Gamma Globin ◽  
Genome Wide ◽  
Hereditary Persistence ◽  
Informative Family ◽  
Gene Switching

Abstract Abstract 5162 We provide additional data on members of the family from Malta with Hereditary Persistence of Fetal Hemoglobin (HPFH) due to KLF1 haplo-insufficiency. The data indicated a possible role of additional loci in the pathway of globin gene control. We showed that KLF1 functions as a master regulator of erythropoiesis and developmental globin gene switching (Borg et al., Nature Genetics doi: 10. 1038/ng.630, 2010), at least partly through BCL11A. Given the phenoytpes of the HPFH heterozygotes, the truncating KLF1 p.K288X was best described as a dominant mutation with variable penetrance; most likely due to interplay with other regulatory factors that we have been seeking. Genome-wide association analysis, in the context of genome-wide expression profiles from cultured Human Erythroid Progenitors (HEPs) of critically informative family members, revealed additional loci of potential interest. The effect of the Hb F inducer Hydroxyurea on the gamma globin profiles of the KLF1- (p.K288X) HPFH HEPs was enhanced compared to the wild type, and 74 loci were differentially expressed. It is anticipated that extensive re-sequencing of these new targets may reveal the extent of the molecular pathways under control of KLF1 in erythropoiesis and globin gene switching, and in particular those that may be targeted for therapeutics in patients. Disclosures: No relevant conflicts of interest to declare.

Human gamma- to beta-globin gene switching using a mini construct in transgenic mice

1992 ◽  
Vol 12 (4) ◽  
pp. 1561-1567
Author(s):  
J A Lloyd ◽  
J M Krakowsky ◽  
S C Crable ◽  
J B Lingrel
Keyword(s):  
Transgenic Mice ◽  
Developmental Regulation ◽  
Globin Gene ◽  
Globin Genes ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Gamma Globin ◽  
High Level ◽  
Gene Switching ◽  
Globin Gene Expression

The developmental regulation of the human globin genes involves a key switch from fetal (gamma-) to adult (beta-) globin gene expression. It is possible to study the mechanism of this switch by expressing the human globin genes in transgenic mice. Previous work has shown that high-level expression of the human globin genes in transgenic mice requires the presence of the locus control region (LCR) upstream of the genes in the beta-globin locus. High-level, correct developmental regulation of beta-globin gene expression in transgenic mice has previously been accomplished only in 30- to 40-kb genomic constructs containing the LCR and multiple genes from the locus. This suggests that either competition for LCR sequences by other globin genes or the presence of intergenic sequences from the beta-globin locus is required to silence the beta-globin gene in embryonic life. The results presented here clearly show that the presence of the gamma-globin gene (3.3 kb) alone is sufficient to down-regulate the beta-globin gene in embryonic transgenic mice made with an LCR-gamma-beta-globin mini construct. The results also show that the gamma-globin gene is down-regulated in adult mice from most transgenic lines made with LCR-gamma-globin constructs not including the beta-globin gene, i.e., that the gamma-globin gene can be autonomously regulated. Evidence presented here suggests that a region 3' of the gamma-globin gene may be important for down-regulation in the adult. The 5'HS2 gamma en beta construct described is a suitable model for further study of the mechanism of human gamma- to beta-globin gene switching in transgenic mice.

scholarly journals Chinese A gamma fetal hemoglobin: C to T substitution at position-196 of the A gamma gene promoter

Blood ◽  
1986 ◽  
Vol 67 (6) ◽  
pp. 1777-1779 ◽  
Author(s):  
R Gelinas ◽  
M Bender ◽  
C Lotshaw ◽  
P Waber ◽  
H Jr Kazazian ◽  
...  
Keyword(s):  
Globin Gene ◽  
Gene Promoter ◽  
Fetal Hemoglobin ◽  
Nucleotide Sequence Analysis ◽  
Globin Genes ◽  
Gamma Globin ◽  
Hemoglobin C ◽  
Hereditary Persistence ◽  
Gene Switching ◽  
Chinese Individual

Abstract The molecular basis for the hereditary persistence of fetal hemoglobin (HPFH) phenotype was studied in a Chinese individual who was heterozygous for a nondeletion form of A gamma-HPFH. Both allelic A gamma-globin genes were isolated by molecular cloning and subjected to nucleotide sequence analysis. One A gamma gene promoter showed a cytosine to thymine transition at position -196, whereas the other promoter was normal. This mutation at position -196 has now ben found in unrelated individuals with the A gamma-HPFH phenotype from Italy, Sardinia, and China, suggesting that it may have arisen independently. The implications of this mutation for models of fetal globin gene switching are discussed.

scholarly journals Chinese A gamma fetal hemoglobin: C to T substitution at position-196 of the A gamma gene promoter

Blood ◽  
1986 ◽  
Vol 67 (6) ◽  
pp. 1777-1779
Author(s):  
R Gelinas ◽  
M Bender ◽  
C Lotshaw ◽  
P Waber ◽  
H Jr Kazazian ◽  
...  
Keyword(s):  
Globin Gene ◽  
Gene Promoter ◽  
Fetal Hemoglobin ◽  
Nucleotide Sequence Analysis ◽  
Globin Genes ◽  
Gamma Globin ◽  
Hemoglobin C ◽  
Hereditary Persistence ◽  
Gene Switching ◽  
Chinese Individual

The molecular basis for the hereditary persistence of fetal hemoglobin (HPFH) phenotype was studied in a Chinese individual who was heterozygous for a nondeletion form of A gamma-HPFH. Both allelic A gamma-globin genes were isolated by molecular cloning and subjected to nucleotide sequence analysis. One A gamma gene promoter showed a cytosine to thymine transition at position -196, whereas the other promoter was normal. This mutation at position -196 has now ben found in unrelated individuals with the A gamma-HPFH phenotype from Italy, Sardinia, and China, suggesting that it may have arisen independently. The implications of this mutation for models of fetal globin gene switching are discussed.

scholarly journals Effects of butyrate and glucocorticoids on gamma- to beta-globin gene switching in somatic cell hybrids.

1995 ◽  
Vol 15 (2) ◽  
pp. 790-795 ◽  
Author(s):  
G Zitnik ◽  
K Peterson ◽  
G Stamatoyannopoulos ◽  
T Papayannopoulou
Keyword(s):  
Globin Gene ◽  
Fetal Hemoglobin ◽  
Cellular Systems ◽  
Erythroid Cell ◽  
Beta Globin ◽  
Erythroid Cells ◽  
Cellular Mechanisms ◽  
Beta Globin Gene ◽  
Gamma Globin ◽  
Gene Switching

Butyrate and its analogs have been shown to induce fetal hemoglobin in humans and primates and in erythroid cell cultures. To obtain insights concerning the cellular mechanisms of butyrate action, we analyzed the effects of butyrate on human globin gene expression in hybrids produced by fusing mouse erythroleukemia cells (MEL) with human fetal erythroid cells (HFE). These hybrids initially express human fetal hemoglobin but subsequently switch to adult globin expression after several weeks in culture. We found that alpha-aminobutyric acid, a butyrate analog which does not induce terminal maturation, strikingly delays the rate of the gamma- to beta-globin gene (gamma-to-beta) switch in the HFE x MEL hybrids. The effect of butyrate on globin expression is transient, with the result that the delay of globin gene switching requires the continuous presence of this compound in culture. Furthermore, butyrate fails to induce fetal hemoglobin expression in hybrids which have switched, suggesting that the effect of this compound on gamma-globin expression is due to inhibition of gamma gene silencing rather than to induction of gamma gene transcription. Since in other cellular systems, glucocorticoids antagonize the action of butyrate, the effect of dexamethasone on the gamma-to-beta switch in HFE x MEL hybrids was examined. Dexamethasone strikingly accelerated the gamma-to-beta switch, and its effect was irreversible. The effects of dexamethasone and butyrate on the gamma-to-beta switch of the HFE x MEL hybrids appear to be codominant. These results indicate that steroids can have a direct effect on globin gene switching in erythroid cells.

scholarly journals Serum factors can modulate the developmental clock of gamma- to beta-globin gene switching in somatic cell hybrids.

1993 ◽  
Vol 13 (8) ◽  
pp. 4844-4851 ◽  
Author(s):  
G Zitnik ◽  
Q Li ◽  
G Stamatoyannopoulos ◽  
T Papayannopoulou
Keyword(s):  
Globin Gene ◽  
Cumulative Number ◽  
Beta Globin ◽  
Serum Factors ◽  
Beta Globin Gene ◽  
Cell Divisions ◽  
Growth Promoting ◽  
Gene Switching ◽  
Mouse Erythroleukemia

The fusion of human fetal erythroid (HFE) cells with mouse erythroleukemia (MEL) cells produces stable synkaryons (HFE x MEL) which can be monitored for extended periods of time in culture. Initially these hybrids express a human fetal globin program (gamma >> beta), but after weeks or months in culture, they switch to an adult pattern of globin expression (beta >> gamma). The rate at which hybrids switch to the adult phenotype is roughly dependent on the gestational age of the fetal erythroid cells used in the fusion, suggesting that the rate of switching in vitro may be determined by a developmental clock type of mechanism, possibly involving the cumulative number of divisions experienced by the human fetal cells. To investigate whether the number or rate of cell divisions postfusion can influence the rate of switching, we monitored the rate of switching in hybrids from independent fusions under growth-promoting (serum-replete) and growth-suppressing (serum-deprived) conditions. We found that hybrids grown under serum-deprived or serumless conditions switched more rapidly to adult globin expression than did their counterparts in serum-replete conditions. Neither the number of cumulative cell divisions nor time in culture per se predicted the rate of switching in vitro. Our data suggest that factors present in serum either retard switching of hybrids by their presence or promote switching by their absence, indicating that globin switching in vitro can be modulated by the environment; however, once switching in HFE x MEL hybrids is complete, serum factors cannot reverse this process.

scholarly journals Loss of allelic heterozygosity at a second locus on chromosome 11 in sporadic Wilms' tumor cells.

1989 ◽  
Vol 9 (4) ◽  
pp. 1799-1803 ◽  
Author(s):  
A E Reeve ◽  
S A Sih ◽  
A M Raizis ◽  
A P Feinberg
Keyword(s):  
Mental Retardation ◽  
Tumor Cells ◽  
Germ Line ◽  
Wilms Tumor ◽  
Globin Gene ◽  
Allelic Loss ◽  
Chromosomal Band ◽  
Chromosome 11 ◽  
Gamma Globin ◽  
Wagr Syndrome

Children with associated Wilms' tumor, aniridia, genitourinary malformations, and mental retardation (WAGR syndrome) frequently have a cytogenetically visible germ line deletion of chromosomal band 11p13. In accordance with the Knudson hypothesis of two-hit carcinogenesis, the absence of this chromosomal band suggests that loss of both alleles of a gene at 11p13 causes Wilms' tumor. Consistent with this model, chromosomes from sporadically occurring Wilms' tumor cells frequently show loss of allelic heterozygosity at polymorphic 11p15 loci, and therefore it has been assumed that allelic loss extends proximally to include 11p13. We report here that in samples from five sporadic Wilms' tumors, allelic loss occurred distal to the WAGR locus on 11p13. In cells from one tumor, mitotic recombination occurred distal to the gamma-globin gene on 11p15.5. Thus, allelic loss in sporadic Wilms' tumor cells may involve a second locus on 11p.

Developmental regulation of β-globin gene switching

Cell ◽  
1988 ◽  
Vol 55 (1) ◽  
pp. 17-26 ◽  
Author(s):  
Ok-Ryun Baik Choi ◽  
James Douglas Engel
Keyword(s):  
Developmental Regulation ◽  
Globin Gene ◽  
Gene Switching
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