Promoter and enhancer elements from the rat elastase I gene function independently of each other and of heterologous enhancers

1987 ◽  
Vol 7 (10) ◽  
pp. 3466-3472
Author(s):  
D M Ornitz ◽  
R E Hammer ◽  
B L Davison ◽  
R L Brinster ◽  
R D Palmiter
Keyword(s):  
Fusion Gene ◽  
Leukemia Virus ◽  
Human Growth ◽  
Cell Types ◽  
Regulatory Elements ◽  
Pancreatic Acinar Cells ◽  
Gene Promoters ◽  
Base Pairs ◽  
Metallothionein Gene ◽  
Rat Elastase

An elastase-human growth hormone (hGH) fusion gene containing 205 base pairs of elastase 5' flanking region is expressed exclusively in pancreatic acinar cells of transgenic mice. This paper shows that the promoter region (-72 to +8) and the enhancer (-205 to -73) function independently of each other. The elastase enhancer can activate the heterologous mouse metallothionein gene and the hGH gene promoters; conversely, enhancers from the thymocyte-specific murine leukemia virus MCF13 and the metal regulatory elements from the metallothionein gene can activate the elastase promoter in a variety of cell types. Combinations of immunoglobulin and elastase enhancers with a heterologous promoter and the hGH gene result in expression in all of the tissues predicted by the sum of each enhancer acting alone. Thus these enhancer elements act independently of each other, suggesting that they do not have silencing activity in cells in which they are normally inactive.

scholarly journals Promoter and enhancer elements from the rat elastase I gene function independently of each other and of heterologous enhancers.

1987 ◽  
Vol 7 (10) ◽  
pp. 3466-3472 ◽  
Author(s):  
D M Ornitz ◽  
R E Hammer ◽  
B L Davison ◽  
R L Brinster ◽  
R D Palmiter
Keyword(s):  
Fusion Gene ◽  
Leukemia Virus ◽  
Human Growth ◽  
Cell Types ◽  
Regulatory Elements ◽  
Pancreatic Acinar Cells ◽  
Gene Promoters ◽  
Base Pairs ◽  
Metallothionein Gene ◽  
Rat Elastase

An elastase-human growth hormone (hGH) fusion gene containing 205 base pairs of elastase 5' flanking region is expressed exclusively in pancreatic acinar cells of transgenic mice. This paper shows that the promoter region (-72 to +8) and the enhancer (-205 to -73) function independently of each other. The elastase enhancer can activate the heterologous mouse metallothionein gene and the hGH gene promoters; conversely, enhancers from the thymocyte-specific murine leukemia virus MCF13 and the metal regulatory elements from the metallothionein gene can activate the elastase promoter in a variety of cell types. Combinations of immunoglobulin and elastase enhancers with a heterologous promoter and the hGH gene result in expression in all of the tissues predicted by the sum of each enhancer acting alone. Thus these enhancer elements act independently of each other, suggesting that they do not have silencing activity in cells in which they are normally inactive.

Upstream G gamma-globin and downstream beta-globin sequences required for stage-specific expression in transgenic mice

1987 ◽  
Vol 7 (11) ◽  
pp. 4024-4029
Author(s):  
M Trudel ◽  
J Magram ◽  
L Bruckner ◽  
F Costantini
Keyword(s):  
Transgenic Mice ◽  
Fusion Gene ◽  
Globin Gene ◽  
Regulatory Elements ◽  
Globin Genes ◽  
Beta Globin ◽  
Erythroid Cells ◽  
Base Pairs ◽  
Beta Globin Gene ◽  
Gamma Globin

The human G gamma-globin and beta-globin genes are expressed in erythroid cells at different stages of human development, and previous studies have shown that the two cloned genes are also expressed in a differential stage-specific manner in transgenic mice. The G gamma-globin gene is expressed only in murine embryonic erythroid cells, while the beta-globin gene is active only at the fetal and adult stages. In this study, we analyzed transgenic mice carrying a series of hybrid genes in which different upstream, intragenic, or downstream sequences were contributed by the beta-globin or G gamma-globin gene. We found that hybrid 5'G gamma/3'beta globin genes containing G gamma-globin sequences upstream from the initiation codon were expressed in embryonic erythroid cells at levels similar to those of an intact G gamma-globin transgene. In contrast, beta-globin upstream sequences were insufficient for expression of 5'beta/3'G gamma hybrid globin genes or a beta-globin-metallothionein fusion gene in adult erythroid cells. However, beta-globin downstream sequences, including 212 base pairs of exon III and 1,900 base pairs of 3'-flanking DNA, were able to activate a 5'G gamma/3'beta hybrid globin gene in fetal and adult erythroid cells. These experiments suggest that positive regulatory elements upstream from the G gamma-globin and downstream from the beta-globin gene are involved in the differential expression of the two genes during development.

Specific expression of an elastase–human growth hormone fusion gene in pancreatic acinar cells of transgenic mice

Nature ◽  
1985 ◽  
Vol 313 (6003) ◽  
pp. 600-602 ◽  
Author(s):  
David M. Ornitz ◽  
Richard D. Palmiter ◽  
Robert E. Hammer ◽  
Ralph L. Brinster ◽  
Galvin H. Swift ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Transgenic Mice ◽  
Human Growth Hormone ◽  
Fusion Gene ◽  
Human Growth ◽  
Acinar Cells ◽  
Pancreatic Acinar Cells ◽  
Specific Expression

scholarly journals Cell type-specific enhancer-promoter connectivity maps in the human brain and disease risk association

2019 ◽  
Author(s):  
Alexi Nott ◽  
Inge R. Holtman ◽  
Nicole G. Coufal ◽  
Johannes C.M. Schlachetzki ◽  
Miao Yu ◽  
...  
Keyword(s):  
Human Brain ◽  
Psychiatric Disorders ◽  
Complex Traits ◽  
Disease Risk ◽  
Cell Types ◽  
Regulatory Elements ◽  
Gene Promoters ◽  
Cell Type ◽  
Risk Variants ◽  
Cell Type Specific

AbstractUnique cell type-specific patterns of activated enhancers can be leveraged to interpret non-coding genetic variation associated with complex traits and diseases such as neurological and psychiatric disorders. Here, we have defined active promoters and enhancers for major cell types of the human brain. Whereas psychiatric disorders were primarily associated with regulatory regions in neurons, idiopathic Alzheimer’s disease (AD) variants were largely confined to microglia enhancers. Interactome maps connecting GWAS variants in cell type-specific enhancers to gene promoters revealed an extended microglia gene network in AD. Deletion of a microglia-specific enhancer harboring AD-risk variants ablated BIN1 expression in microglia but not in neurons or astrocytes. These findings revise and expand the genes likely to be influenced by non-coding variants in AD and suggest the probable brain cell types in which they function.One Sentence SummaryIdentification of cell type-specific regulatory elements in the human brain enables interpretation of non-coding GWAS risk variants.

scholarly journals An atlas of silencer elements for the human and mouse genomes

2018 ◽  
Author(s):  
Naresh Doni Jayavelu ◽  
Ajay Jajodia ◽  
Arpit Mishra ◽  
R. David Hawkins
Keyword(s):  
Cell Types ◽  
Regulatory Elements ◽  
Mouse Cell ◽  
Support Vector ◽  
General Strategy ◽  
Gene Promoters ◽  
Genome Wide ◽  
A Genome ◽  
Silencer Elements ◽  
Human And Mouse

ABSTRACTThe study of gene regulation is dominated by a focus on the control of gene activation or controlling an increase in the level of expression. Just as critical is the process of gene repression or silencing. Chromatin signatures have allowed for the global mapping of enhancer cis-regulatory elements, however, the identification of silencer elements by computational or experimental approaches in a genome-wide manner are lacking. We present a simple but powerful computational approach to identify putative silencers genome-wide. We used a series of consortia data to predict silencers in over 100 human and mouse cell or tissue types. We performed several analyses to determine if these elements exhibited characteristics expected of a silencers. Motif enrichment analyses on putative silencers determined that motifs belonging to known transcriptional repressors are enriched, as well as overlapping known transcription repressor binding sites. Leveraging promoter capture HiC data from several human and mouse cell types, we found that over 50% of putative silencer elements are interacting with gene promoters having very low to no expression. Next, to validate our silencer predictions, we quantified silencer activity using massively parallel reporter assays (MPRAs) on 7500 selected elements in K562 cells. We trained a support vector machine model classifier on MPRA data and used it to refine potential silencers in other cell types. We also show that similar to enhancer elements, silencer elements are enriched in disease-associated variants. Our results suggest a general strategy for genome-wide identification and characterization of silencer elements.

Building a metal-responsive promoter with synthetic regulatory elements

1985 ◽  
Vol 5 (6) ◽  
pp. 1480-1489
Author(s):  
P F Searle ◽  
G W Stuart ◽  
R D Palmiter
Keyword(s):  
Thymidine Kinase ◽  
Promoter Region ◽  
Fusion Gene ◽  
Regulatory Elements ◽  
Regulatory Sequence ◽  
Thymidine Kinase Gene ◽  
Coding Region ◽  
Base Pairs ◽  
Promoter Elements ◽  
Kinase Gene

A fusion gene consisting of the promoter region from the mouse metallothionein-I gene joined to the coding region of the herpes simplex virus thymidine kinase gene is efficiently regulated by zinc in a transient assay when transfected into baby hamster kidney cells. Analysis of similar plasmids in which the metallothionein-I promoter region was mutated indicated the presence of multiple metal regulatory elements (MREs) between -176 and -44 base pairs from the cap site. To further investigate the function of MREs, we inserted a synthetic DNA fragment containing the sequence of MRE-a (the element between -55 and -44 base pairs) into the nonresponsive promoter of the thymidine kinase gene in various positions and configurations. Little or no induction by zinc was observed with single insertions of the regulatory sequence, whereas many different constructions having two copies of MRE-a were inducible. The precise position of the two MREs relative to each other or to the thymidine kinase promoter elements had a relatively small effect on the efficiency of induction, but the inducibility could be further increased by the introduction of more MRE-a sequences. MRE-a can function synergistically with the thymidine kinase distal promoter elements, but in the presence of the TATA box alone it functions as a positive, zinc-dependent promoter element.

Glucocorticoid regulation of human growth hormone expression in transgenic mice and transiently transfected cells

1989 ◽  
Vol 122 (1) ◽  
pp. 49-60 ◽  
Author(s):  
R. F Selden ◽  
J. S. Yun ◽  
D. D. Moore ◽  
M. E. Rowe ◽  
M. A. Malia ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Transgenic Mice ◽  
Human Growth Hormone ◽  
Dna Sequences ◽  
Fusion Gene ◽  
Human Growth ◽  
Regulatory Elements ◽  
Growth Hormone Gene ◽  
L Cells ◽  
Gene Mrna

ABSTRACT A mouse metallothionein-I/human growth hormone fusion gene was microinjected into fertilized mouse eggs, the embryos were implanted into pseudopregnant foster mothers, and the offspring analysed. Five of twenty-six mice born after one series of injections contained from one to eight copies of the fusion gene stably integrated into their genomes and had human growth hormone in their serum. When several of these transgenic mice and transgenic offspring were treated with glucocorticoids, serum growth hormone levels were elevated from 1·5- to 6·3-fold. A fourfold induction in fusion gene mRNA in the liver of one of the five mice was also observed after treatment with glucocorticoids. When the fusion gene was transiently transfected into mouse L cells, dexamethasone caused a three- to fourfold induction of fusion gene mRNA and secreted human growth hormone. A deletion analysis of regulatory elements required for inducibility in L cells shows that DNA sequences responsible for the observed inductions are located within the transcribed region of the human growth hormone gene. However, a previously described glucocorticoid receptor binding site in the first intron of the gene is not required for response to the hormone. Journal of Endocrinology (1989) 122, 49–60

scholarly journals Building a metal-responsive promoter with synthetic regulatory elements.

1985 ◽  
Vol 5 (6) ◽  
pp. 1480-1489 ◽  
Author(s):  
P F Searle ◽  
G W Stuart ◽  
R D Palmiter
Keyword(s):  
Thymidine Kinase ◽  
Promoter Region ◽  
Fusion Gene ◽  
Regulatory Elements ◽  
Regulatory Sequence ◽  
Thymidine Kinase Gene ◽  
Coding Region ◽  
Base Pairs ◽  
Promoter Elements ◽  
Kinase Gene

A fusion gene consisting of the promoter region from the mouse metallothionein-I gene joined to the coding region of the herpes simplex virus thymidine kinase gene is efficiently regulated by zinc in a transient assay when transfected into baby hamster kidney cells. Analysis of similar plasmids in which the metallothionein-I promoter region was mutated indicated the presence of multiple metal regulatory elements (MREs) between -176 and -44 base pairs from the cap site. To further investigate the function of MREs, we inserted a synthetic DNA fragment containing the sequence of MRE-a (the element between -55 and -44 base pairs) into the nonresponsive promoter of the thymidine kinase gene in various positions and configurations. Little or no induction by zinc was observed with single insertions of the regulatory sequence, whereas many different constructions having two copies of MRE-a were inducible. The precise position of the two MREs relative to each other or to the thymidine kinase promoter elements had a relatively small effect on the efficiency of induction, but the inducibility could be further increased by the introduction of more MRE-a sequences. MRE-a can function synergistically with the thymidine kinase distal promoter elements, but in the presence of the TATA box alone it functions as a positive, zinc-dependent promoter element.

scholarly journals Upstream G gamma-globin and downstream beta-globin sequences required for stage-specific expression in transgenic mice.

1987 ◽  
Vol 7 (11) ◽  
pp. 4024-4029 ◽  
Author(s):  
M Trudel ◽  
J Magram ◽  
L Bruckner ◽  
F Costantini
Keyword(s):  
Transgenic Mice ◽  
Fusion Gene ◽  
Globin Gene ◽  
Regulatory Elements ◽  
Globin Genes ◽  
Beta Globin ◽  
Erythroid Cells ◽  
Base Pairs ◽  
Beta Globin Gene ◽  
Gamma Globin

The human G gamma-globin and beta-globin genes are expressed in erythroid cells at different stages of human development, and previous studies have shown that the two cloned genes are also expressed in a differential stage-specific manner in transgenic mice. The G gamma-globin gene is expressed only in murine embryonic erythroid cells, while the beta-globin gene is active only at the fetal and adult stages. In this study, we analyzed transgenic mice carrying a series of hybrid genes in which different upstream, intragenic, or downstream sequences were contributed by the beta-globin or G gamma-globin gene. We found that hybrid 5'G gamma/3'beta globin genes containing G gamma-globin sequences upstream from the initiation codon were expressed in embryonic erythroid cells at levels similar to those of an intact G gamma-globin transgene. In contrast, beta-globin upstream sequences were insufficient for expression of 5'beta/3'G gamma hybrid globin genes or a beta-globin-metallothionein fusion gene in adult erythroid cells. However, beta-globin downstream sequences, including 212 base pairs of exon III and 1,900 base pairs of 3'-flanking DNA, were able to activate a 5'G gamma/3'beta hybrid globin gene in fetal and adult erythroid cells. These experiments suggest that positive regulatory elements upstream from the G gamma-globin and downstream from the beta-globin gene are involved in the differential expression of the two genes during development.

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