Specific expression of a foreign β-globin gene in erythroid cells of transgenic mice

Chada, Kiran; Magram, Jeanne; Raphael, Kathryn A.; Radice, Glenn L.; Lacy, Elizabeth; Costantini, Frank

doi:10.1038/314377a0

Cited by 226 publications

(104 citation statements)

References 29 publications

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“…Surprisingly, all 5' deletion clones still maintained an appropriate up-regulation as the cells differentiated into myofibers except when combined with pBR322 sequences, which drastically inhibit transcription. The nature of this inhibitory effect is unknown, but it has been observed in a number of transgenic mouse experiments (5,53). One possibility for the inhibition of expression is that the high G+C content of plasmid sequences may produce structural alterations that inhibit interactions of the promoter with regulatory factors on RNA polymerase.…”

Section: Discussionmentioning

confidence: 99%

Complex regulation of the muscle-specific contractile protein (troponin I) gene.

1987

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A cloned quail troponin I contractile protein gene, stably transfected into a mouse myogenic cell line, exhibits appropriate developmental activation and quantitative expression during myoblast differentiation. Deletion mutagenesis analyses reveal that the troponin I gene has two distinct cis regulatory elements required for its developmental expression, as measured by mRNA accumulation and nuclear runoff transcription assays. One element in the 5' flanking region is required for maximum quantitative expression, and a second larger regulatory element (1.5 kilobases) within the first intron is responsible for differentiation-specific transcription.The upstream region is highly sensitive to negative repression by interaction with pBR322 sequences. The larger intragenic region retains some activity when moved to the 5' and 3' flanking regions and when inverted but is maximally active in its native intragenic site. The concerted activities of these two regulatory regions produce a 100-to 200-fold transcriptional activation during myoblast differentiation. The conserved 5' exon-intron organization of troponin I and other contractile protein genes suggests a possible mechanism by which intragenic control elements coordinate contractile protein gene regulation during skeletal myogenesis.

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Section: Discussionmentioning

confidence: 99%

Complex regulation of the muscle-specific contractile protein (troponin I) gene.

1987

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“…The presence of the LCR is an absolute requirement for the high level expression of the genes in the cluster [2,3]. The linkage of a human ]3-globin transgene to the LCR results in high level, erythroid specific expression in mice [4], in sharp contrast to constructs lacking LCR sequences which show a low, variable expression level [5][6][7][8]. The most characteristic feature of the LCR is its ability to override position effects when it is stably integrated into chromatin.…”

Section: Introductionmentioning

confidence: 99%

Transcriptional activation by hypersensitive site three of the human β-globin locus control region in murine erythroleukemia cells

Pruzina¹,

Antoniou²,

Hurst³

et al. 1994

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expres

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In this paper we describe a complete deletional analysis of hypersensitive site three (HS3) of the human /3-globin Locus Control Region (LCR). The previously defined core fragment consists of 6 footprinted regions, with multiple binding sites for the erythroid-specific factor GATA-1 and G-rich motifs that can interact with ubiquitous factors such as Spl and TEF-2. We show in this paper that the 5' half of this fragment is the most important for activity in murine erythroleukemia (MEL) cells. A fragment containing footprints 1-4 can stimulate transcription of a linked human /3-globin gene to levels of about 40% of that obtained with footprints 1-6. Constructs containing either footprints 1-3 or 3-6 cannot be distinguished from the/3-globin gene alone. We further show that binding sites for the erythroid-specific factor NF-E2 can co-operatively interact with parts of the HS3 core fragment, and that HS3 requires elements upstream from -103 in the human /3-globin promoter for full activity. The importance of these results is discussed in the context of the regulation of the genes in the human /3-globin cluster.

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“…Furthermore, LaFlamme et al (27) transfected chimeric globin genes involving the exchange of the IVS2 between the human ,B-and b-globin genes into MEL cells and demonstrated that the IVS2 of human ,B-globin is necessary, but not sufficient, for inducible expression of the gene in MEL cells. In transgenic mice, the human ,B-globin gene is regulated as a mouse adult globin gene and expressed in both fetal and adult stages (9,29,48) and the human -y-globin gene is regulated as an embryonic globin gene (8). Chimeric lyl$-globin genes (joined at the BamHI site at the 3' end of exon 2) in transgenic mice are expressed as both embryonic and adult ,-globin genes (26), further implicating sequences downstream of exon 2 in the regulation of the ,B-globin gene.…”

mentioning

confidence: 99%

Detection of two tissue-specific DNA-binding proteins with affinity for sites in the mouse beta-globin intervening sequence 2.

Galson¹,

Housman²

1988

Mol. Cell. Biol.

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To identify proteins from uninduced murine erythroleukemia nuclear extracts which specifically bind to sequences from the DNase I-hypersensitive region within the mouse I-globin intervening sequence 2 (IVS2), a gel electrophoretic mobility shift assay was used. Two distinct sequence-specific binding proteins were detected. The specific binding sites for these factors were delineated by both DNase I protection footprinting and methylation interference. Factor Bl bound specifically to two homologous sites, B1-A and B1-B, approximately 100 base pairs apart within the IVS2 and on opposite strands. These two regions could interact with factor B1 independently. Factor B1 was limited to cells of hematopoietic lineages. Factor B2 bound to a site approximately 5 base pairs away from the B1-A site and was limited to cells of the erythroid lineage. The limited tissue distribution of these factors and the locations of their binding sites suggest that one or both of these factors may be involved in the formation of the tissue-specific DNase I-hypersensitive site in the IVS2 of the mouse I-globin gene.

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Specific expression of a foreign β-globin gene in erythroid cells of transgenic mice

Cited by 226 publications

References 29 publications

Complex regulation of the muscle-specific contractile protein (troponin I) gene.

Complex regulation of the muscle-specific contractile protein (troponin I) gene.

Transcriptional activation by hypersensitive site three of the human β-globin locus control region in murine erythroleukemia cells

Detection of two tissue-specific DNA-binding proteins with affinity for sites in the mouse beta-globin intervening sequence 2.

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