Maria Giuseppina Marini
Researcher
Area of interest:
Maria Giuseppina Marini is Researcher at the IRGB CNR in Monserrato (CA), Italy.
She graduated in Biological Sciences at the University of Cagliari, Italy, in 1991, obtained a specialization in Genetics and Molecular Biology at the University of Bologna, Italy, in 1997
and a doctorate at the University of Cagliari, Italy, in 2006.
Field of research: Molecular Pathology of thalassemias and other genetic diseases, gene regulation, identification of the etio-pathogenic basis of susceptibility to pathogen resistance.
Project leader of CRP_ CRP-60653 Grant Regione Sardegna
Most significant publications:
2021
Mingoia, M; Caria, C A; Ye, L; Asunis, I; Marongiu, M F; Manunza, L; Sollaino, M C; Wang, J; Cabriolu, A; Kurita, R; Nakamura, Y; Cucca, F; Kan, Y W; Marini, M G; Moi, P
Induction of therapeutic levels of HbF in genome-edited primary β0 39-thalassaemia haematopoietic stem and progenitor cells Journal Article
In: Br J Haematol, 192 (2), pp. 395–404, 2021.
@article{pmid33216968,
title = {Induction of therapeutic levels of HbF in genome-edited primary β0 39-thalassaemia haematopoietic stem and progenitor cells},
author = {M Mingoia and C A Caria and L Ye and I Asunis and M F Marongiu and L Manunza and M C Sollaino and J Wang and A Cabriolu and R Kurita and Y Nakamura and F Cucca and Y W Kan and M G Marini and P Moi},
year = {2021},
date = {2021-01-01},
journal = {Br J Haematol},
volume = {192},
number = {2},
pages = {395--404},
abstract = {Hereditary persistence of fetal haemoglobin (HPFH) is the major modifier of the clinical severity of β-thalassaemia. The homozygous mutation c.-196 C>T in the Aγ-globin (HBG1) promoter, which causes Sardinian δβ0 -thalassaemia, is able to completely rescue the β-major thalassaemia phenotype caused by the β0 39-thalassaemia mutation, ensuring high levels of fetal haemoglobin synthesis during adulthood. Here, we describe a CRISPR/Cas9 genome-editing approach, combined with the non-homologous end joining (NHEJ) pathway repair, aimed at reproducing the effects of this naturally occurring HPFH mutation in both HBG promoters. After selecting the most efficient guide RNA in K562 cells, we edited the HBG promoters in human umbilical cord blood-derived erythroid progenitor 2 cells (HUDEP-2) and in haematopoietic stem and progenitor cells (HSPCs) from β0 -thalassaemia patients to assess the therapeutic potential of HbF induction. Our results indicate that small deletions targeting the -196-promoter region restore high levels of fetal haemoglobin (HbF) synthesis in all cell types tested. In pools of HSPCs derived from homozygous β0 39-thalassaemia patients, a 20% editing determined a parallel 20% increase of HbF compared to unedited pools. These results suggest that editing the region of HBG promoters around the -196 position has the potential to induce therapeutic levels of HbF in patients with most types of β-thalassaemia irrespective of the β-globin gene (HBB) mutations.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Hereditary persistence of fetal haemoglobin (HPFH) is the major modifier of the clinical severity of β-thalassaemia. The homozygous mutation c.-196 C>T in the Aγ-globin (HBG1) promoter, which causes Sardinian δβ0 -thalassaemia, is able to completely rescue the β-major thalassaemia phenotype caused by the β0 39-thalassaemia mutation, ensuring high levels of fetal haemoglobin synthesis during adulthood. Here, we describe a CRISPR/Cas9 genome-editing approach, combined with the non-homologous end joining (NHEJ) pathway repair, aimed at reproducing the effects of this naturally occurring HPFH mutation in both HBG promoters. After selecting the most efficient guide RNA in K562 cells, we edited the HBG promoters in human umbilical cord blood-derived erythroid progenitor 2 cells (HUDEP-2) and in haematopoietic stem and progenitor cells (HSPCs) from β0 -thalassaemia patients to assess the therapeutic potential of HbF induction. Our results indicate that small deletions targeting the -196-promoter region restore high levels of fetal haemoglobin (HbF) synthesis in all cell types tested. In pools of HSPCs derived from homozygous β0 39-thalassaemia patients, a 20% editing determined a parallel 20% increase of HbF compared to unedited pools. These results suggest that editing the region of HBG promoters around the -196 position has the potential to induce therapeutic levels of HbF in patients with most types of β-thalassaemia irrespective of the β-globin gene (HBB) mutations.
2010
Marini, M. G.; Porcu, L.; Asunis, I.; Loi, M. G.; Ristaldi, M. S.; Porcu, S.; Ikuta, T.; Cao, A.; Moi, P.
Regulation of the human ĦBA genes by KLF4 in erythroid cell lines Journal Article
In: Br J Haematol, 149 (5), pp. 748–758, 2010.
@article{pmid20331458,
title = {Regulation of the human ĦBA genes by KLF4 in erythroid cell lines},
author = { M. G. Marini and L. Porcu and I. Asunis and M. G. Loi and M. S. Ristaldi and S. Porcu and T. Ikuta and A. Cao and P. Moi},
year = {2010},
date = {2010-06-01},
journal = {Br J Haematol},
volume = {149},
number = {5},
pages = {748--758},
abstract = {KLF1/EKLF and related Krueppel-like factors (KLFs) are variably implicated in the regulation of the HBB-like globin genes. Prompted by the observation that four KLF sites are distributed in the human alpha-globin gene (HBA) promoter, we investigated if KLFs could also act to modulate the expression of the HBA genes. Among the KLFs tested, only KLF4/GKLF bound specifically to three out of four alpha-globin KLF sites. The occupancy of the same sites by KLF4 in vivo was confirmed by chromatin immunoprecipitation assays with KLF4-specific antibodies. In luciferase reporter assays in MEL cells, high levels of the wild type HBA promoter, but not mutated promoters bearing point mutations that disrupted KLF4-DNA binding, were transactivated by over-expression of KLF4. In K562 cells, induced KLF4 expression with a Tet-off regulated cassette stimulated the expression of the endogenous HBA genes. In a complementary assay in the same cell line, knocking down KLF4 with lentiviral delivered sh-RNAs caused a parallel decrease in the transcription of the HBA genes. All experiments combined support a regulatory role of KLF4 in the control of HBA gene expression.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
KLF1/EKLF and related Krueppel-like factors (KLFs) are variably implicated in the regulation of the HBB-like globin genes. Prompted by the observation that four KLF sites are distributed in the human alpha-globin gene (HBA) promoter, we investigated if KLFs could also act to modulate the expression of the HBA genes. Among the KLFs tested, only KLF4/GKLF bound specifically to three out of four alpha-globin KLF sites. The occupancy of the same sites by KLF4 in vivo was confirmed by chromatin immunoprecipitation assays with KLF4-specific antibodies. In luciferase reporter assays in MEL cells, high levels of the wild type HBA promoter, but not mutated promoters bearing point mutations that disrupted KLF4-DNA binding, were transactivated by over-expression of KLF4. In K562 cells, induced KLF4 expression with a Tet-off regulated cassette stimulated the expression of the endogenous HBA genes. In a complementary assay in the same cell line, knocking down KLF4 with lentiviral delivered sh-RNAs caused a parallel decrease in the transcription of the HBA genes. All experiments combined support a regulatory role of KLF4 in the control of HBA gene expression.
2004
Marini, M. G.; Asunis, I.; Porcu, L.; Salgo, M. G.; Loi, M. G.; Brucchietti, A.; Cao, A.; Moi, P.
The distal beta-globin CACCC box is required for maximal stimulation of the beta-globin gene by EKLF Journal Article
In: Br J Haematol, 127 (1), pp. 114–117, 2004.
@article{pmid15384985,
title = {The distal beta-globin CACCC box is required for maximal stimulation of the beta-globin gene by EKLF},
author = { M. G. Marini and I. Asunis and L. Porcu and M. G. Salgo and M. G. Loi and A. Brucchietti and A. Cao and P. Moi},
year = {2004},
date = {2004-01-01},
journal = {Br J Haematol},
volume = {127},
number = {1},
pages = {114--117},
abstract = {The transcription factor erythroid Kruppel-like factor (EKLF) specifically activates the beta-globin gene by interacting with the proximal beta-globin CACCC box, a known hot spot for thalassaemia mutations. This study investigated whether EKLF could also bind to, and activate from, the distal CACCC, which is a rare site of thalassaemia mutations. Using band shift and transient expression analysis with wild type, single and double CACCC mutants, we established that the distal CACCC box is weakly bound by EKLF, but, when mutated, significantly impairs EKLF-dependent beta-globin stimulation. Thus, EKLF requires both CACCC boxes to maximally stimulate the beta-globin gene.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The transcription factor erythroid Kruppel-like factor (EKLF) specifically activates the beta-globin gene by interacting with the proximal beta-globin CACCC box, a known hot spot for thalassaemia mutations. This study investigated whether EKLF could also bind to, and activate from, the distal CACCC, which is a rare site of thalassaemia mutations. Using band shift and transient expression analysis with wild type, single and double CACCC mutants, we established that the distal CACCC box is weakly bound by EKLF, but, when mutated, significantly impairs EKLF-dependent beta-globin stimulation. Thus, EKLF requires both CACCC boxes to maximally stimulate the beta-globin gene.
2002
Marini, M. G.; Asunis, I.; Chan, K.; Chan, J. Y.; Kan, Y. W.; Porcu, L.; Cao, A.; Moi, P.
Cloning MafF by recognition site screening with the NFE2 tandem repeat of ĦS2: analysis of its role in globin and GCSl genes regulation Journal Article
In: Blood Cells Mol Dis, 29 (2), pp. 145–158, 2002.
@article{pmid12490281,
title = {Cloning MafF by recognition site screening with the NFE2 tandem repeat of ĦS2: analysis of its role in globin and GCSl genes regulation},
author = { M. G. Marini and I. Asunis and K. Chan and J. Y. Chan and Y. W. Kan and L. Porcu and A. Cao and P. Moi},
year = {2002},
date = {2002-01-01},
journal = {Blood Cells Mol Dis},
volume = {29},
number = {2},
pages = {145--158},
abstract = {The erythroid-specific enhancer within hypersensitivity site 2 (HS2) of the human beta-globin locus control region is required for high level globin gene expression. We used an oligonucleotide of the NF-E2 tandem repeat, within HS2, as recognition site probe to screen a K562 cDNA library for interacting transcription factors. A 2.3 kb full length cDNA encoding the b-zip transcription factor MafF was isolated. MafF can form both homodimers and high affinity heterodimers with Nrf1, Nrf2 and Nf-E2, three members of the CNC-bZip family. Despite obvious structural similarities with the other small Maf proteins, MafF differs in its tissue distribution and its inability to repress transcription when overexpressed as homodimer. In fact, in different cell lines and on different promoters (gamma-globin, beta-globin and glutamylcysteine synthetase genes) the MafF homodimers do not appreciably affect transcription of target promoters, whereas MafF/CNC member heterodimers act as weak transcriptional activators. Even though MafF was cloned using probes derived from the globin LCR, it is in the context of the GCSl promoter and in combination with Jun that MafF shows a rather distinct and specific regulatory role. These observations suggest that a complex network of small Maf and CNC-AP1 protein interactions might be involved in regulating transcription in diverse tissues or developmental stages.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The erythroid-specific enhancer within hypersensitivity site 2 (HS2) of the human beta-globin locus control region is required for high level globin gene expression. We used an oligonucleotide of the NF-E2 tandem repeat, within HS2, as recognition site probe to screen a K562 cDNA library for interacting transcription factors. A 2.3 kb full length cDNA encoding the b-zip transcription factor MafF was isolated. MafF can form both homodimers and high affinity heterodimers with Nrf1, Nrf2 and Nf-E2, three members of the CNC-bZip family. Despite obvious structural similarities with the other small Maf proteins, MafF differs in its tissue distribution and its inability to repress transcription when overexpressed as homodimer. In fact, in different cell lines and on different promoters (gamma-globin, beta-globin and glutamylcysteine synthetase genes) the MafF homodimers do not appreciably affect transcription of target promoters, whereas MafF/CNC member heterodimers act as weak transcriptional activators. Even though MafF was cloned using probes derived from the globin LCR, it is in the context of the GCSl promoter and in combination with Jun that MafF shows a rather distinct and specific regulatory role. These observations suggest that a complex network of small Maf and CNC-AP1 protein interactions might be involved in regulating transcription in diverse tissues or developmental stages.
1997
Marini, M. G.; Chan, K.; Casula, L.; Kan, Y. W.; Cao, A.; Moi, P.
hMAF, a small human transcription factor that heterodimerizes specifically with Nrf1 and Nrf2 Journal Article
In: J Biol Chem, 272 (26), pp. 16490–16497, 1997.
@article{pmid9195958,
title = {hMAF, a small human transcription factor that heterodimerizes specifically with Nrf1 and Nrf2},
author = { M. G. Marini and K. Chan and L. Casula and Y. W. Kan and A. Cao and P. Moi},
year = {1997},
date = {1997-06-01},
journal = {J Biol Chem},
volume = {272},
number = {26},
pages = {16490--16497},
abstract = {A 1.6-kilobase pair full-length cDNA encoding a transcription factor homologous to the Maf family of proteins was isolated by screening a K562 cDNA library with the NFE2 tandem repeat probe derived from the globin locus control region. The protein, which was designated hMAF, contains a basic DNA binding domain and an extended leucine zipper but lacks any recognizable activation domain. Expressed in vitro, the hMAF protein is able to homodimerize in solution and band-shift the NFE2 tandem repeat probe. In addition to homodimers, hMAF can also form high affinity heterodimers with two members of the NFE2/CNC-bZip family (Nrf1 and Nrf2) but not with a third family member, p45-NFE2. Although hMAF/hMAF homodimers and hMAF/Nrf1 and hMAF/Nrf2 heterodimers bind to the same NFE2 site, they exert functionally opposite effects on the activity of a linked gamma-globin gene. In fact, whereas all hMAF/CNC-bZip heterodimers stimulate the activity of a gamma-promoter reporter construct in K562 cells, the association into homodimers that is induced by overexpressing hMAF inhibits the activity of the same construct. Thus variations in the expression of hMAF may account for the modulation in the activity of the genes that bear NFE2 recognition sites.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
A 1.6-kilobase pair full-length cDNA encoding a transcription factor homologous to the Maf family of proteins was isolated by screening a K562 cDNA library with the NFE2 tandem repeat probe derived from the globin locus control region. The protein, which was designated hMAF, contains a basic DNA binding domain and an extended leucine zipper but lacks any recognizable activation domain. Expressed in vitro, the hMAF protein is able to homodimerize in solution and band-shift the NFE2 tandem repeat probe. In addition to homodimers, hMAF can also form high affinity heterodimers with two members of the NFE2/CNC-bZip family (Nrf1 and Nrf2) but not with a third family member, p45-NFE2. Although hMAF/hMAF homodimers and hMAF/Nrf1 and hMAF/Nrf2 heterodimers bind to the same NFE2 site, they exert functionally opposite effects on the activity of a linked gamma-globin gene. In fact, whereas all hMAF/CNC-bZip heterodimers stimulate the activity of a gamma-promoter reporter construct in K562 cells, the association into homodimers that is induced by overexpressing hMAF inhibits the activity of the same construct. Thus variations in the expression of hMAF may account for the modulation in the activity of the genes that bear NFE2 recognition sites.
Monserrato
070 6754598
ORCID profile: https://orcid.org/0000-0003-0371-9398