The rat elastase I regulatory element is an enhancer that directs correct cell specificity and developmental onset of expression in transgenic mice

1987 ◽  
Vol 7 (8) ◽  
pp. 2956-2967
Author(s):  
R E Hammer ◽  
G H Swift ◽  
D M Ornitz ◽  
C J Quaife ◽  
R D Palmiter ◽  
...  
Keyword(s):  
Fusion Gene ◽  
Regulatory Element ◽  
Regulatory Region ◽  
Human Growth ◽  
Transgenic Animals ◽  
Growth Hormone Gene ◽  
Flanking Sequence ◽  
Transcriptional Enhancer ◽  
Independent Manner ◽  
Specific Expression

A total of 134 base pairs of the 5' flanking sequence of the elastase I gene is sufficient and necessary to direct expression of the passive human growth hormone gene (hGH) to the exocrine pancreas. We demonstrate that this elastase I regulatory region contains a transcriptional enhancer which directs acinar cell-specific expression in transgenic animals. The elastase I enhancer specifies correct expression of the linked hGH gene in an orientation- and position-independent manner and can activate a heterologous promoter. The enhancer also directs the appropriate temporal activation of the hGH gene in the developing pancreas. Transcription is initiated correctly for the elastase I or hGH promoter, and the transcripts are correctly processed regardless of the enhancer position within or outside the fusion gene. The elastase I enhancer generates coincident DNase I-hypersensitive sites in pancreatic chromatin when moved 3 kilobases upstream or within the first intron of the hGH gene and when associated with the hGH promoter.

scholarly journals The rat elastase I regulatory element is an enhancer that directs correct cell specificity and developmental onset of expression in transgenic mice.

1987 ◽  
Vol 7 (8) ◽  
pp. 2956-2967 ◽  
Author(s):  
R E Hammer ◽  
G H Swift ◽  
D M Ornitz ◽  
C J Quaife ◽  
R D Palmiter ◽  
...  
Keyword(s):  
Fusion Gene ◽  
Regulatory Element ◽  
Regulatory Region ◽  
Human Growth ◽  
Transgenic Animals ◽  
Growth Hormone Gene ◽  
Flanking Sequence ◽  
Transcriptional Enhancer ◽  
Independent Manner ◽  
Specific Expression

A total of 134 base pairs of the 5' flanking sequence of the elastase I gene is sufficient and necessary to direct expression of the passive human growth hormone gene (hGH) to the exocrine pancreas. We demonstrate that this elastase I regulatory region contains a transcriptional enhancer which directs acinar cell-specific expression in transgenic animals. The elastase I enhancer specifies correct expression of the linked hGH gene in an orientation- and position-independent manner and can activate a heterologous promoter. The enhancer also directs the appropriate temporal activation of the hGH gene in the developing pancreas. Transcription is initiated correctly for the elastase I or hGH promoter, and the transcripts are correctly processed regardless of the enhancer position within or outside the fusion gene. The elastase I enhancer generates coincident DNase I-hypersensitive sites in pancreatic chromatin when moved 3 kilobases upstream or within the first intron of the hGH gene and when associated with the hGH promoter.

Binding of a factor to an enhancer element responsible for the tissue-specific expression of the chicken alpha A-crystallin gene

Development ◽  
1991 ◽  
Vol 113 (2) ◽  
pp. 539-550 ◽  
Author(s):  
I. Matsuo ◽  
M. Kitamura ◽  
K. Okazaki ◽  
K. Yasuda
Keyword(s):  
Gene Expression ◽  
Dna Sequences ◽  
Nuclear Protein ◽  
Fusion Gene ◽  
Regulatory Region ◽  
Binding Activity ◽  
Flanking Sequence ◽  
Specific Expression ◽  
Enhancer Element

We have characterized a regulatory region of the chicken alpha A-crystallin gene using transfection assays, which revealed that a 84 base pair element (−162 to −79) in the 5′ flanking sequence is necessary and sufficient for lens-specific expression. A multimer of this element functions as lens-specific enhancer and synergistically activates transcription from chicken alpha A-crystallin or beta-actin basal promoters fused to the CAT gene. In vivo competition experiments demonstrated that DNA sequences containing the 84 bp element reduced alpha A-crystallin-CAT fusion gene expression. A nuclear factor present exclusively in lens cells binds to the 84 bp element in the region between positions −165 and −140. Southwestern blot analysis showed that 61,000 Mr (61 × 10(3) Mr) lens nuclear protein exhibited DNA-binding activity specific to the 84 bp element. Our data suggested that the 61 × 10(3) Mr nuclear protein, and the 84 bp element that it interacts with, may be involved in regulating the alpha A-crystallin gene expression in vivo.

Meiotic expression of human ornithine transcarbamylase in the testes of transgenic mice

1988 ◽  
Vol 8 (4) ◽  
pp. 1821-1825
Author(s):  
K A Kelley ◽  
J W Chamberlain ◽  
J A Nolan ◽  
A L Horwich ◽  
F Kalousek ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Fusion Gene ◽  
Regulatory Region ◽  
Transgenic Animals ◽  
Ornithine Transcarbamylase ◽  
Regulatory Sequences ◽  
Base Pairs ◽  
Protein Coding ◽  
Wide Range ◽  
Flanking Sequences

In an attempt to use mouse metallothionein-I (mMT-I) regulatory sequences to direct expression of human ornithine transcarbamylase in the liver of transgenic animals, fusion genes joining either 1.6 kilobases or 185 base pairs of the mMT-I regulatory region to the human ornithine transcarbamylase protein-coding sequence were used to produce transgenic mice. In mice carrying the fusion gene with 1.6 kilobases of the mMT-I 5'-flanking sequences, transgene expression was observed in a wide range of tissues, but, unexpectedly, expression in liver was never observed. Surprisingly, in mice carrying the fusion gene regulated by only 185 base pairs of the mMT-I 5'-flanking sequences, the transgene was expressed exclusively in male germ cells during the tetraploid, pachytene stage of meiosis.

Introduction of a porcine growth hormone fusion gene into transgenic pigs promotes growth

1988 ◽  
Vol 90 (2) ◽  
pp. 295-300
Author(s):  
P.D. Vize ◽  
A.E. Michalska ◽  
R. Ashman ◽  
B. Lloyd ◽  
B.A. Stone ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Fusion Gene ◽  
Transgenic Animals ◽  
Growth Hormone Gene ◽  
Gene Sequences ◽  
Transgenic Pigs ◽  
Gene Construct ◽  
Metallothionein Promoter

Six transgenic pigs have been produced by microinjecting a human metallothionein promoter/porcine growth hormone gene construct into the pronuclei of fertilized eggs which were transferred to synchronized recipient sows. The resulting transgenic animals contained between 0.5 and 15 copies of the gene construct per cell, and at least one of the animals expressed the introduced gene and grew at an increased rate compared to both transgenic and non-transgenic littermates. Some of the transgenic animals that did not appear to grow at increased rates were found to contain rearranged gene sequences. Two of the transgenic pigs have been shown to pass on the introduced genes to their offspring.

Targeting gene expression to the head: the Drosophila orthodenticle gene is a direct target of the Bicoid morphogen

Development ◽  
1998 ◽  
Vol 125 (21) ◽  
pp. 4185-4193 ◽  
Author(s):  
Q. Gao ◽  
R. Finkelstein
Keyword(s):  
Gene Expression ◽  
Target Genes ◽  
Regulatory Element ◽  
Regulatory Region ◽  
Repeated Sequence ◽  
Drosophila Embryo ◽  
Sequence Motif ◽  
Specific Expression ◽  
Gap Gene

The Bicoid (Bcd) morphogen establishes the head and thorax of the Drosophila embryo. Bcd activates the transcription of identified target genes in the thoracic segments, but its mechanism of action in the head remains poorly understood. It has been proposed that Bcd directly activates the cephalic gap genes, which are the first zygotic genes to be expressed in the head primordium. It has also been suggested that the affinity of Bcd-binding sites in the promoters of Bcd target genes determines the posterior extent of their expression (the Gene X model). However, both these hypotheses remain untested. Here, we show that a small regulatory region upstream of the cephalic gap gene orthodenticle (otd) is sufficient to recapitulate early otd expression in the head primordium. This region contains two control elements, each capable of driving otd-like expression. The first element has consensus Bcd target sites that bind Bcd in vitro and are necessary for head-specific expression. As predicted by the Gene X model, this element has a relatively low affinity for Bcd. Surprisingly, the second regulatory element has no Bcd sites. Instead, it contains a repeated sequence motif similar to a regulatory element found in the promoters of otd-related genes in vertebrates. Our study is the first demonstration that a cephalic gap gene is directly regulated by Bcd. However, it also shows that zygotic gene expression can be targeted to the head primordium without direct Bcd regulation.

A rat gastrin-human gastrin chimeric transgene directs antral G cell-specific expression in transgenic mice

1995 ◽  
Vol 268 (6) ◽  
pp. G1025-G1036 ◽  
Author(s):  
T. C. Wang ◽  
M. W. Babyatsky ◽  
P. S. Oates ◽  
Z. Zhang ◽  
L. Tillotson ◽  
...  
Keyword(s):  
Germ Line ◽  
Peptide Yy ◽  
Human Growth ◽  
Growth Hormone Gene ◽  
Specific Expression ◽  
Cis Acting ◽  
G Cells ◽  
Human Gastrin ◽  
Immunohistochemical Studies

Gastrin gene expression in the gastrointestinal tract is under both developmental and spatial regulation. In the mature animal, gastrin, an important regulator of parietal acid secretion, is expressed primarily in G cells of the antrum. To determine whether specific promoter elements can direct expression to the gastric antrum in vivo, 450 nucleotides of the proximal rat gastrin promoter were cloned and used to construct a rat gastrin-human gastrin reporter chimeric transgene, which was injected into the mouse germ line. Northern blot analysis, in situ hybridization, and double-label immunocytochemistry studies demonstrated expression of the transgene specifically in antral G cells. Low levels of transgene expression were observed in the ileum and colon, where immunohistochemical studies demonstrated colocalization in enteroendocrine cells expressing peptide YY. The same 450-nucleotide rat gastrin promoter, when joined to the human growth hormone gene, did not result in antral expression. Similarly, a human gastrin-human gastrin reporter transgene also did not achieve antral expression, although it did express in the liver. These results suggest that cis-acting elements present in both the basal 450-nucleotide rat gastrin promoter and the intragenic sequences of the human gastrin gene are necessary to direct expression of a transgene specifically to antral G cells.

scholarly journals Role of the 5′ enhancer of the glutamine synthetase gene in its organ-specific expression

1997 ◽  
Vol 323 (3) ◽  
pp. 611-619 ◽  
Author(s):  
Heleen LIE-VENEMA ◽  
Piet A. J. DE BOER ◽  
Antoon F. M. MOORMAN ◽  
Wouter H. LAMERS
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Gastrointestinal Tract ◽  
Glutamine Synthetase ◽  
Regulatory Element ◽  
Regulatory Region ◽  
Upstream Regulatory Region ◽  
Specific Expression ◽  
Organ Specific ◽  
Ribonuclease Protection

In mammals, glutamine synthetase (GS) is expressed in a large number of organs, but the precise regulation of its expression is still obscure. Therefore a detailed analysis of the activity of the upstream regulatory element of the GS gene in the transcriptional regulation of its expression was carried out in transgenic mice carrying the chloramphenicol acetyltransferase (CAT) gene under the control of the upstream regulatory region of the GS gene. CAT and GS mRNA expression were compared in liver, epididymis, lung, adipocytes, testis, kidney, skeletal muscle and gastrointestinal tract, both quantitatively by ribonuclease-protection analysis and topographically by in situ hybridization. It was found that the upstream regulatory region is active with respect both to the level and to the topography of GS gene expression in liver, epididymis, gastrointestinal tract (stomach, small intestine and colon) and skeletal muscle. On the other hand, in the kidney, brain, adipocytes, spleen, lung and testis, GS gene expression is not or only partly regulated by the 5´ enhancer. A second enhancer, identified within the first intron, may regulate GS expression in the latter organs. Furthermore, CAT expression in the brain did not co-localize with that of GS, showing that the 5´ regulatory region of the GS gene does not direct its expression to the astrocytes.

scholarly journals Differentiation-responsive elements in the 5' region of the mouse tissue plasminogen activator gene confer two-stage regulation by retinoic acid and cyclic AMP in teratocarcinoma cells.

1989 ◽  
Vol 9 (4) ◽  
pp. 1691-1704 ◽  
Author(s):  
R J Rickles ◽  
A L Darrow ◽  
S Strickland
Keyword(s):  
Retinoic Acid ◽  
Cyclic Amp ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Regulatory Element ◽  
Mouse Tissue ◽  
Flanking Sequence ◽  
Two Stage ◽  
Specific Expression ◽  
Tissue Plasminogen

F9 cells induced to differentiate with retinoic acid (RA) increase transcription of the tissue plasminogen activator (t-PA) gene. Further treatment of these cells with cyclic AMP (cAMP) results in an additional stimulation of t-PA gene transcription. To investigate the mechanism of this two-stage regulation, 4 kilobase pairs (kbp) of 5'-flanking sequence from the murine t-PA gene was isolated. Two major start sites for transcription were found, neither of which depended on a classical TATA motif for correct initiation. By using transient transfection assays, it was determined that 4-kbp of flanking sequence could confer on reporter genes the same two-stage differentiation-specific expression as was observed for the endogenous t-PA gene. Deletion analyses of this 4-kbp fragment showed that 190 bp of flanking sequence was sufficient to bestow the same degree of two-stage regulation on reporter gene constructs. Within this region of DNA, sequence analysis revealed a possible cAMP regulatory element, a CTF/NF-1 recognition sequence, two potential Sp1 sites, and five potential binding sites for transcription factor AP-2. The deletion experiments, coupled with the positions of these potential cis-acting elements, suggest that multiple transcription factors, including those that bind to cAMP regulatory element, CTF/NF-1, Sp1, and AP-2 sites, may be involved in regulation of the t-PA gene during F9 cell differentiation.

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