Tissue-specific and hormonally regulated expression of a rat alpha 2u globulin gene in transgenic mice

1987 ◽  
Vol 7 (10) ◽  
pp. 3749-3758
Author(s):  
V da C Soares ◽  
R M Gubits ◽  
P Feigelson ◽  
F Costantini
Keyword(s):  
Transgenic Mice ◽  
Gene Family ◽  
Hormonal Regulation ◽  
Germ Line ◽  
Regulatory Elements ◽  
Preputial Gland ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Globulin Gene

To investigate the tissue-specific and hormonal regulation of the rat alpha 2u globulin gene family, we introduced one cloned member of the gene family into the mouse germ line and studied its expression in the resulting transgenic mice. Alpha 2u globulingene 207 was microinjected on a 7-kilobase DNA fragment, and four transgenic lines were analyzed. The transgene was expressed at very high levels, specifically in the liver and the preputial gland of adult male mice. The expression in male liver was first detected at puberty, and no expression was detected in female transgenic mice. This pattern of expression is similar to the expression of endogenous alpha 2u globulin genes in the rat but differs from the expression of the homologous mouse major urinary protein (MUP) gene family in that MUPs are synthesized in female liver and not in the male preputial gland. We conclude that these differences between rat alpha 2u globulin and mouse MUP gene expression are due to evolutionary differences in cis-acting regulatory elements. The expression of the alpha 2u globulin transgene in the liver was abolished by castration and fully restored after testosterone replacement. The expression could also be induced in the livers of female mice by treatment with either testosterone or dexamethasone, following ovariectomy and adrenalectomy. Therefore, the cis-acting elements responsible for regulation by these two hormones, as well as those responsible for tissue-specific expression, are closely linked to the alpha 2u globulin gene.

scholarly journals Tissue-specific and hormonally regulated expression of a rat alpha 2u globulin gene in transgenic mice.

1987 ◽  
Vol 7 (10) ◽  
pp. 3749-3758 ◽  
Author(s):  
V da C Soares ◽  
R M Gubits ◽  
P Feigelson ◽  
F Costantini
Keyword(s):  
Transgenic Mice ◽  
Gene Family ◽  
Hormonal Regulation ◽  
Germ Line ◽  
Regulatory Elements ◽  
Preputial Gland ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Globulin Gene

To investigate the tissue-specific and hormonal regulation of the rat alpha 2u globulin gene family, we introduced one cloned member of the gene family into the mouse germ line and studied its expression in the resulting transgenic mice. Alpha 2u globulingene 207 was microinjected on a 7-kilobase DNA fragment, and four transgenic lines were analyzed. The transgene was expressed at very high levels, specifically in the liver and the preputial gland of adult male mice. The expression in male liver was first detected at puberty, and no expression was detected in female transgenic mice. This pattern of expression is similar to the expression of endogenous alpha 2u globulin genes in the rat but differs from the expression of the homologous mouse major urinary protein (MUP) gene family in that MUPs are synthesized in female liver and not in the male preputial gland. We conclude that these differences between rat alpha 2u globulin and mouse MUP gene expression are due to evolutionary differences in cis-acting regulatory elements. The expression of the alpha 2u globulin transgene in the liver was abolished by castration and fully restored after testosterone replacement. The expression could also be induced in the livers of female mice by treatment with either testosterone or dexamethasone, following ovariectomy and adrenalectomy. Therefore, the cis-acting elements responsible for regulation by these two hormones, as well as those responsible for tissue-specific expression, are closely linked to the alpha 2u globulin gene.

scholarly journals Tissue-specific regulation of the expression of rat kallikrein gene family members by thyroid hormone

1990 ◽  
Vol 267 (3) ◽  
pp. 745-750 ◽  
Author(s):  
J A Clements ◽  
B A Matheson ◽  
J E Funder
Keyword(s):  
Gene Family ◽  
Hormonal Regulation ◽  
Female Rats ◽  
Hormonal Status ◽  
Mrna Levels ◽  
Specific Expression ◽  
Male And Female ◽  
Tissue Specific ◽  
Male And Female Rats ◽  
Specific Regulation

We have altered the thyroid hormonal status of both male and female rats and examined the expression of six functional members of the rat kallikrein gene family (PS, S1, S2, S3, K1 and P1) in the submandibular gland (SMG), kidney, prostate, testis and anterior pituitary gland (AP) of these animals. On Northern-blot analysis with gene-specific oligonucleotide probes, the steady-state mRNA levels of S1, S2, S3, K1 and P1 were all dramatically altered in the SMG of male and female rats treated with propylthiouracil (PTU; 100 mg/litre of drinking water) or thyroxine (T4; 10 micrograms/100 mg body wt.) for 3 weeks. The SMG mRNA levels of these five genes were all lowered (30-90%) in hypothyroid (PTU-treated) male and female rats and elevated (1.4-4-fold, male; 1.5-11-fold, female) in the hyperthyroid (T4-treated) and PTU/T4-treated animals. In contrast, PS (true kallikrein) mRNA levels in the male or female SMG or kidney were essentially unchanged. K1 mRNA levels in the kidney were considerably less responsive to thyroid status than those in the SMG. Changes in S3 and P1 mRNA levels in the prostate were also variable, but essentially unaffected by these treatments. AP PS mRNA levels were also unaffected by changes in thyroid-hormonal status, as were levels of a novel P1-like mRNA in the testis. In summary, these studies demonstrate that the same kallikrein gene family member(s) may be differentially regulated by thyroid hormones in the rat SMG, kidney, prostate and pituitary, and thus further extend the concept of tissue-specific expression and hormonal regulation of the kallikrein gene family in the rat.

scholarly journals Tissue-specific, developmental, hormonal, and dietary regulation of rat phosphoenolpyruvate carboxykinase-human growth hormone fusion genes in transgenic mice.

1992 ◽  
Vol 12 (3) ◽  
pp. 1007-1020 ◽  
Author(s):  
M K Short ◽  
D E Clouthier ◽  
I M Schaefer ◽  
R E Hammer ◽  
M A Magnuson ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Transgenic Mice ◽  
Human Growth Hormone ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Human Growth ◽  
Fusion Genes ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Specific Manner

The cytosolic phosphoenolpyruvate carboxykinase (PEPCK) gene is expressed in multiple tissues and is regulated in a complex tissue-specific manner. To map the cis-acting DNA elements that direct this tissue-specific expression, we made transgenic mice containing truncated PEPCK-human growth hormone (hGH) fusion genes. The transgenes contained PEPCK promoter fragments with 5' endpoints at -2088, -888, -600, -402, and -207 bp, while the 3' endpoint was at +69 bp. Immunohistochemical analysis showed that the -2088 transgene was expressed in the correct cell types (hepatocytes, proximal tubular epithelium of the kidney, villar epithelium of the small intestine, epithelium of the colon, smooth muscle of the vagina and lungs, ductal epithelium of the sublingual gland, and white and brown adipocytes). Solution hybridization of hGH mRNA expressed from the transgenes indicated that white and brown fat-specific elements are located distally (-2088 to -888 bp) and that liver-, gut-, and kidney-specific elements are located proximally (-600 to +69 bp). However, elements outside of the region tested are necessary for the correct developmental pattern and level of PEPCK expression in kidney. Both the -2088 and -402 transgenes responded in a tissue-specific manner to dietary stimuli, and the -2088 transgene responded to glucocorticoid stimuli. Thus, different tissues utilize distinct cell-specific cis-acting elements to direct and regulate the PEPCK gene.

Identification of cis sequences controlling efficient position-independent tissue-specific expression of human major histocompatibility complex class I genes in transgenic mice

1991 ◽  
Vol 11 (7) ◽  
pp. 3564-3572 ◽  
Author(s):  
J W Chamberlain ◽  
H A Vasavada ◽  
S Ganguly ◽  
S M Weissman
Keyword(s):  
Major Histocompatibility Complex ◽  
Transgenic Mice ◽  
Major Histocompatibility Complex Class ◽  
Class I ◽  
Complex Class ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Tissue Specific Expression ◽  
Histocompatibility Complex

We previously reported that genomic major histocompatibility complex class I human leukocyte antigen (HLA)-B7 gene constructs with as little as 0.66 kb of 5'- and 2.0 kb of 3'-flanking DNA were expressed efficiently and appropriately in transgenic mice. To identify and characterize the relevant cis-acting regulatory elements in more detail, we have generated and analyzed a series of transgenic mice carrying native HLA-B7 genes with further 5' truncations or intronic deletions and hybrid constructs linking the 5'-flanking region of B7 to a reporter gene. We were unable to detect a specific requirement for sequence information within introns 2 to 7 for either appropriate constitutive or inducible class I expression in adult animals. The results revealed the presence of cis-acting regulatory sequences between -0.075 kb and -0.66 kb involved in driving efficient copy number-dependent constitutive and gamma interferon-enhanced tissue-specific expression. The region from -0.11 to -0.66 kb is also sufficient to prevent integration site-specific "position effects," because in its absence HLA-B7 expression is frequently detected at significant levels at inappropriate sites. Conserved sequence elements homologous to the H-2 class I regulatory element, or enhancer A, and the interferon response sequence are located between about -151 and -228 bp of the B7 gene. Our results also indicate the existence of sequences downstream of -0.11 kb which can influence the pattern of tissue-specific expression of the HLA-B7 gene and the ability of this gene to respond to gamma interferon.

scholarly journals Ksp-cadherin gene promoter. II. Kidney-specific activity in transgenic mice

1999 ◽  
Vol 277 (4) ◽  
pp. F599-F610 ◽  
Author(s):  
Peter Igarashi ◽  
Cooduvalli S. Shashikant ◽  
R. Brent Thomson ◽  
Dilys A. Whyte ◽  
Shuxian Liu-Chen ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Transgenic Mice ◽  
Collecting Duct ◽  
Gene Promoter ◽  
Regulatory Elements ◽  
Developmental Expression ◽  
Tubular Epithelial Cells ◽  
Specific Expression ◽  
Tissue Specific

Kidney-specific cadherin (Ksp-cadherin, cadherin 16) is a tissue-specific member of the cadherin superfamily that is expressed exclusively in the basolateral membrane of tubular epithelial cells in the kidney. To determine the basis for tissue-specific expression of Ksp-cadherin in vivo, we evaluated the activity of the promoter in transgenic mice. Transgenic mice containing 3.3 kb of the mouse Ksp-cadherin promoter and an Escherichia coli lacZ reporter gene were generated by pronuclear microinjection. Assays of β-galactosidase enzyme activity showed that the transgene was expressed exclusively in the kidney in both adult and developing mice. Within the kidney, the transgene was expressed in a subset of renal tubular epithelial cells that endogenously expressed Ksp-cadherin and that were identified as collecting ducts by colabeling with Dolichos biflorus agglutinin. In the developing metanephros, expression of the transgene in the branching ureteric bud correlated with the developmental expression of Ksp-cadherin. Identical patterns of expression were observed in multiple founder mice, indicating that kidney specificity was independent of transgene integration site. However, heterocellular expression was observed consistent with repeat-induced gene silencing. We conclude that the Ksp-cadherin gene promoter directs kidney-specific expression in vivo. Regulatory elements that are sufficient to recapitulate the tissue- and differentiation-specific expression of Ksp-cadherin in the renal collecting duct are located within 3.3 kb upstream to the transcriptional start site.

A modified pod specific promoter for high level heterologous expression of genes in legumes

2019 ◽  
Author(s):  
Aravind Kumar Konda ◽  
Pallavi Singh ◽  
Khela Ram Soren ◽  
Narendra Pratap Singh
Keyword(s):  
Gene Expression ◽  
Regulatory Elements ◽  
Specific Gene ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Inducible Promoters ◽  
Expression Of Genes ◽  
Enhanced Expression ◽  
High Level

Promoters are cis-acting regulatory elements that are usually present upstream to the coding sequences and determine the gene expression. Deployment of tissue specific and inducible promoters are constantly increasing for development of successful and stable multiple transgenic plants. To this end, as a strategy for enhanced expression of cis or transgenes, promoter engineering of the native msg promoter from soya bean has been carried out for executing pod specific expression of genes. Cis regulatory elements such as 5’UTR and poly (A) tract have been incorporated for imparting mRNA stability and translational enhancement to generate the modified 1.285 Kb pod specific promoter. Further to attain transcriptional enhancement the modified promoter has been cloned to generate Bi-directional Duplex Promoters (BDDP). The engineered msg promoter gene constructs can be deployed for high level tissue specific gene expression of cis/trans genes along with chosen terminator in chickpea. soybean and other legumes as well.

scholarly journals Genomic Regions that Mediate Placental Cell-Specific and Developmental Regulation of Human Cyp19 (Aromatase) Gene Expression in Transgenic Mice

Endocrinology ◽  
2005 ◽  
Vol 146 (5) ◽  
pp. 2481-2488 ◽  
Author(s):  
Amrita Kamat ◽  
Margaret E. Smith ◽  
John M. Shelton ◽  
James A. Richardson ◽  
Carole R. Mendelson
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Developmental Regulation ◽  
Giant Cells ◽  
Regulatory Elements ◽  
Deletion Mapping ◽  
Flanking Sequence ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cyp19 Gene

Abstract The human aromatase (hCYP19) gene is controlled by tissue-specific promoters that lie upstream of tissue-specific first exons. Placenta-specific exon I.1 lies approximately 100,000 bp upstream of exon II. Previously, we observed that genomic sequences within 501 bp upstream of exon I.1 mediate placenta-specific expression. In the present study, transgenic mice were created carrying hCYP19I.1−246:hGH/hGX, hCYP19I.1−201:hGH, and hCYP19I.1−125:hGH fusion genes to further delineate 5′-flanking sequences within 501 bp of exon I.1 that are required to mediate placenta-specific hCYP19 gene expression. As little as 246 bp of hCYP19 exon I.1 5′-flanking sequence was sufficient to direct placenta-specific expression in transgenic mice. By contrast, transgenes containing 201 or 125 bp of exon I.1 5′-flanking DNA were not expressed in mouse placenta. Furthermore, hCYP19I.1−246:hGX transgene expression was developmentally regulated; expression was observed as early as embryonic d 7.5 (E7.5) in several cells of the trophoblast ectoderm, on E8.5 in some trophoblast giant cells, and by E9.5 in giant cells and the labyrinthine layer. By contrast, expression of the hCYP19I.1−501:hGH transgene was first observed on E10.5 and was restricted to the labyrinthine layer, which is most analogous to the human syncytiotrophoblast. This suggests the presence of regulatory elements between −501 and −246 bp that may bind inhibitory transcription factors expressed in giant cells. These findings from transgenic experiments together with deletion mapping studies using transfected human placental cells indicate that the concerted interaction of strong placenta-specific enhancers and silencers within this 501-bp region mediate labyrinthine and syncytiotrophoblast-specific CYP19 gene expression.

scholarly journals GENETIC LOCALIZATION AND ACTION OF REGULATORY GENES AND ELEMENTS FOR TISSUE-SPECIFIC EXPRESSION OF α-AMYLASE IN DROSOPHILA MELANOGASTER

Genetics ◽  
1986 ◽  
Vol 114 (4) ◽  
pp. 1131-1145
Author(s):  
A J Klarenberg ◽  
A J S Visser ◽  
M F M Willemse ◽  
W Scharloo
Keyword(s):  
Drosophila Melanogaster ◽  
Regulatory Gene ◽  
Regulatory Elements ◽  
Specific Expression ◽  
Tissue Specific ◽  
Cis Acting ◽  
Single Adults ◽  
Posterior Midgut ◽  
Trans Regulation ◽  
Anterior Midgut

ABSTRACT Regulation of tissue-specific α-amylase (Amy) expression in Drosophila melanogaster was investigated with a newly developed method that detects the distribution of α-amylase allozymes in midguts of single adults or third-instar larvae. Trans regulation was found for α-amylase production in the posterior midgut (PMG) of adults, whereas cis regulation was demonstrated for the larval midgut. Independent regulation of components of the duplicated Amy locus was found in larvae. Recombination between the components of the Amy locus did not result in separation of the putative, very closely linked (less than 0.1 cM) cis-acting regulatory elements for α-amylase expression in the anterior midgut (AMG) of larvae. The expression of one of the components of the duplicated Amy locus in the AMG of larvae was influenced by a regulatory gene that was mapped at 2-79.1. α-Amylase expression in the adult PMG was controlled by a trans-acting regulatory gene localized at 2-79.0, in agreement with the data of Abraham and Doane.

scholarly journals Tissue-specific expression and promoter analyses of the human tissue kallikrein gene in transgenic mice

1997 ◽  
Vol 325 (1) ◽  
pp. 111-116 ◽  
Author(s):  
William XIONG ◽  
Jing WANG ◽  
Lee CHAO ◽  
Julie CHAO
Keyword(s):  
Transgenic Mice ◽  
Human Tissue ◽  
Promoter Region ◽  
Translational Regulation ◽  
Regulatory Elements ◽  
Tissue Kallikrein ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression ◽  
Specific Expression Pattern

The expression of the tissue kallikrein gene is tissue-specific and exhibits a complex pattern of transcriptional and post-translational regulation. Information concerning the mechanism of its tissue-specific expression has been limited owing to the lack of suitable cell lines for the expression study. We approached this problem by introducing human tissue kallikrein gene constructs into mouse embryos, creating transgenic lines carrying its coding sequence with varying lengths of the promoter region. One construct (PHK) contained 801 bp in the 5′-flanking region and two deletion constructs contained either 302 bp (D300) or 202 bp (D200) of the promoter region. The expression of human tissue kallikrein in these transgenic mice was monitored by Northern blot, reverse transcriptase–PCR followed by Southern blot, and radioimmunoassay. In all three lines, human tissue kallikrein was expressed predominantly in the pancreas and at lower levels in other tissues, including salivary gland, kidney and spleen. This pattern was similar to that of tissue kallikrein expression in human tissues. The D300 line has higher levels of transgene expression than the D200 and PHK lines. The results indicate that the 202 bp segment immediately upstream of the translation starting site is sufficient to direct a tissue-specific expression pattern of the human tissue kallikrein gene, and that regulatory elements might exist between -801 and -202.

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