scholarly journals Minimal DNA sequences that control the cell lineage-specific expression of the pro alpha 2(I) collagen promoter in transgenic mice.

1992 ◽  
Vol 119 (5) ◽  
pp. 1361-1370 ◽  
Author(s):  
K Niederreither ◽  
R N D'Souza ◽  
B de Crombrugghe
Keyword(s):  
Transgenic Mice ◽  
Upstream Sequence ◽  
Collagen Gene ◽  
Galactosidase Activity ◽  
Tgf Beta ◽  
Specific Expression ◽  
Cultured Fibroblasts ◽  
Tissue Specific ◽  
Collagen Promoter ◽  
Beta Galactosidase

The pattern of expression of the pro alpha 2(I) collagen gene is highly tissue specific in adult mice and shows its strongest expression in bones, tendons, and skin. Transgenic mice were generated harboring promoter fragments of the mouse pro alpha 2(I) collagen gene linked to the Escherichia coli beta-galactosidase or firefly luciferase genes to examine the activity of these promoters during development. A region of the mouse pro alpha 2(I) collagen promoter between -2,000 and +54 exhibited a pattern of beta-galactosidase activity during embryonic development that corresponded to the expression pattern of the endogenous pro alpha 2(I) collagen gene as determined by in situ hybridization. A similar pattern of activity was also observed with much smaller promoter fragments containing either 500 or 350 bp of upstream sequence relative to the start of transcription. Embryonic regions expressing high levels of beta-galactosidase activity included the bulbus arteriosus, valves of the developing heart, sclerotomes, meninges, limb buds, connective tissue fascia between muscle fibers, osteoblasts in newly formed bones, fibroblasts in tendons, periosteum, dermis, and peritoneal membranes. The pattern of beta-galactosidase activity was similar and included within the extracellular immunohistochemical localization pattern of transforming growth factor-beta 1 (TGF-beta 1). The -315(-)-284 region of the pro alpha 2(I) collagen promoter was previously shown to mediate the stimulatory effects of TGF-beta 1 on the pro alpha 2(I) collagen promoter in DNA transfection experiments with cultured fibroblasts. A construct containing this sequence tandemly repeated 5' to a very short alpha 2(I) collagen promoter (-40(-)+54) showed preferential activity in tail and skin of 4-wk-old transgenic mice. Except for low expression of the transgene in bone, this pattern mimics the expression of the endogenous pro alpha 2(I) collagen gene. We propose the hypothesis that the tissue-specific expression of the pro alpha 2(I) collagen gene during embryogenesis is controlled by both TGF-beta 1 and cell-specific transcription factors; one of these could interact directly or indirectly with either the -315(-)-284 or the -40(-)+54 segment.

A 182 bp fragment of the mouse pro alpha 1(II) collagen gene is sufficient to direct chondrocyte expression in transgenic mice

1995 ◽  
Vol 108 (12) ◽  
pp. 3677-3684 ◽  
Author(s):  
G. Zhou ◽  
S. Garofalo ◽  
K. Mukhopadhyay ◽  
V. Lefebvre ◽  
C.N. Smith ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Reporter Gene ◽  
Type Ii Collagen ◽  
Mouse Embryos ◽  
Collagen Gene ◽  
Specific Expression ◽  
Intact Mouse ◽  
Endogenous Gene ◽  
Intron 1 ◽  
Beta Galactosidase

Type II collagen is a major chondrocyte-specific component of the cartilage extracellular matrix and it represents a typical differentiation marker of mature chondrocytes. In order to delineate cis-acting elements of the mouse pro alpha 1(II) collagen gene that control chondrocyte-specific expression in intact mouse embryos, we generated transgenic mice harboring chimeric constructions in which varying lengths of the promoter and intron 1 sequences were linked to a beta-galactosidase reporter gene. A construction containing a 3,000 bp promoter and a 3,020 bp intron 1 fragment directed high levels of beta-galactosidase expression specifically to chondrocytes. Expression of the transgene coincided with the temporal expression of the endogenous gene at all stages of embryonic development. Successive deletions of intron 1 delineated a 182 bp fragment which targeted beta-galactosidase expression to chondrocytes with the same specificity as the larger intron 1 fragment. Transgenic mice harboring a 309 bp Col2a1 promoter lacking intron 1 tester sequences showed no beta-galactosidase expression in chondrocytes. Reduction of the 182 bp fragment to a 73 bp subfragment surrounding a decamer sequence previously reported to be involved in chondrocyte specificity, resulted in loss of transgene expression in chondrocytes. When the Col2a1 promoter was replaced with a minimal beta-globin promoter, the 182 bp intron 1 sequence was still able to target expression of the transgene to chondrocytes. We conclude that a 182 bp intron 1 DNA segment of the mouse Col2a1 gene contains the necessary information to confer high-level, temporally correct, chondrocyte expression on a reporter gene in intact mouse embryos and that Col2a1 promoter sequences are dispensable for chondrocyte expression.

scholarly journals Developmental and tissue-specific expression directed by the alpha 2 type I collagen promoter in transgenic mice.

1986 ◽  
Vol 83 (3) ◽  
pp. 725-729 ◽  
Author(s):  
J. S. Khillan ◽  
A. Schmidt ◽  
P. A. Overbeek ◽  
B. de Crombrugghe ◽  
H. Westphal
Keyword(s):  
Transgenic Mice ◽  
Type I Collagen ◽  
Type I ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression ◽  
Collagen Promoter

scholarly journals An upstream regulatory region mediates high-level, tissue-specific expression of the human alpha 1(I) collagen gene in transgenic mice.

1991 ◽  
Vol 11 (4) ◽  
pp. 2066-2074 ◽  
Author(s):  
J L Slack ◽  
D J Liska ◽  
P Bornstein
Keyword(s):  
Transgenic Mice ◽  
Regulatory Region ◽  
Collagen Gene ◽  
Flanking Sequence ◽  
Specific Expression ◽  
Tissue Specific ◽  
Endogenous Gene ◽  
Tissue Specific Expression ◽  
First Intron ◽  
High Level

Studies in vitro have not adequately resolved the role of intronic and upstream elements in regulating expression of the alpha 1(I) collagen gene. To address this issue, we generated 12 separate lines of transgenic mice with alpha 1(I) collagen-human growth hormone (hGH) constructs containing different amounts of 5'-flanking sequence, with or without most of the first intron. Transgenes driven by 2.3 kb of alpha 1(I) 5'-flanking sequence, whether or not they contained the first intron, were expressed at a high level and in a tissue-specific manner in seven out of seven independent lines of transgenic mice. In most tissues, the transgene was expressed at levels approaching that of the endogenous alpha 1(I) gene and was regulated identically with the endogenous gene as animals aged. However, in lung, expression of the transgene was anomalously high, and in muscle, expression was lower than that of the endogenous gene, suggesting that in these tissues other regions of the gene may participate in directing appropriate expression. Five lines of mice were generated containing transgenes driven by 0.44 kb of alpha 1(I) 5'-flanking sequence (with or without the first intron), and expression was detected in four out of five of these lines. The level of expression of the 0.44-kb constructs in the major collagen-producing tissues was 15- to 500-fold lower than that observed with the longer 2.3-kb promoter. While transgenes containing the 0.44-kb promoter and the first intron retained a modest degree of tissue-specific expression, those without the first intron lacked tissue specificity and were poorly expressed in all tissues except lung.(ABSTRACT TRUNCATED AT 250 WORDS)

Tissue-Specific Expression of Promoter Regions of the alphar1(VI) Collagen Gene in Cell Cultures and Transgenic Mice

1997 ◽  
Vol 247 (1) ◽  
pp. 200-208 ◽  
Author(s):  
Paola Braghetta ◽  
Paola Vitale ◽  
Stefano Piccolo ◽  
Paolo Bonaldo ◽  
Carla Fabbro ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Cell Cultures ◽  
Collagen Gene ◽  
Promoter Regions ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression

An upstream regulatory region mediates high-level, tissue-specific expression of the human alpha 1(I) collagen gene in transgenic mice

1991 ◽  
Vol 11 (4) ◽  
pp. 2066-2074
Author(s):  
J L Slack ◽  
D J Liska ◽  
P Bornstein
Keyword(s):  
Transgenic Mice ◽  
Regulatory Region ◽  
Collagen Gene ◽  
Flanking Sequence ◽  
Specific Expression ◽  
Tissue Specific ◽  
Endogenous Gene ◽  
Tissue Specific Expression ◽  
First Intron ◽  
High Level

Studies in vitro have not adequately resolved the role of intronic and upstream elements in regulating expression of the alpha 1(I) collagen gene. To address this issue, we generated 12 separate lines of transgenic mice with alpha 1(I) collagen-human growth hormone (hGH) constructs containing different amounts of 5'-flanking sequence, with or without most of the first intron. Transgenes driven by 2.3 kb of alpha 1(I) 5'-flanking sequence, whether or not they contained the first intron, were expressed at a high level and in a tissue-specific manner in seven out of seven independent lines of transgenic mice. In most tissues, the transgene was expressed at levels approaching that of the endogenous alpha 1(I) gene and was regulated identically with the endogenous gene as animals aged. However, in lung, expression of the transgene was anomalously high, and in muscle, expression was lower than that of the endogenous gene, suggesting that in these tissues other regions of the gene may participate in directing appropriate expression. Five lines of mice were generated containing transgenes driven by 0.44 kb of alpha 1(I) 5'-flanking sequence (with or without the first intron), and expression was detected in four out of five of these lines. The level of expression of the 0.44-kb constructs in the major collagen-producing tissues was 15- to 500-fold lower than that observed with the longer 2.3-kb promoter. While transgenes containing the 0.44-kb promoter and the first intron retained a modest degree of tissue-specific expression, those without the first intron lacked tissue specificity and were poorly expressed in all tissues except lung.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals The immediate upstream sequence of the mouse Ret gene controls tissue-specific expression in transgenic mice

2006 ◽  
Author(s):  
Paola Zordan ◽  
Sara Tavella ◽  
Antonella Brizzolara ◽  
Roberta Biticchi ◽  
Isabella Ceccherini ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Upstream Sequence ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression ◽  
Ret Gene

scholarly journals Cell-specific expression of alpha 1(I) collagen-hGH minigenes in transgenic mice.

1994 ◽  
Vol 125 (3) ◽  
pp. 695-704 ◽  
Author(s):  
D J Liska ◽  
M J Reed ◽  
E H Sage ◽  
P Bornstein
Keyword(s):  
Skeletal Muscle ◽  
Transgenic Mice ◽  
Cultured Cells ◽  
Dermal Fibroblasts ◽  
Collagen Gene ◽  
Flanking Sequence ◽  
Regulation Of Expression ◽  
Specific Expression ◽  
Cultured Fibroblasts ◽  
First Intron

Sequences within the first intron of the alpha 1(I) collagen gene have been implicated in the regulation of expression of alpha 1(I) collagen-reporter gene constructs in cultured cells. However, the physiological significance of these intronic elements has not been established. We have used in situ hybridization to examine whether a cell-specific pattern of expression of human alpha 1(I) collagen-human growth hormone minigenes exists in transgenic mice. Our results indicate that transgenes which contained 2,300 bp of promoter/5' flanking sequence and an intact first intron were well expressed by fibroblasts in dermis and fascia, whereas transgenes lacking the intronic sequence, +292 to +1440, were not expressed in dermis and poorly expressed in fascia. Analysis of transgene expression in cultured fibroblasts obtained from dermal explants of transgenic animals confirmed the requirement for these intronic sequences in the regulation of the alpha 1(I) collagen gene. In contrast, transgenes with or without the intronic deletion were expressed equally well in tendon and bone, in a manner comparable to the endogenous mouse alpha 1(I) collagen gene, and expression of neither transgene was detected in skeletal muscle or perichondrium. These data support a model in which cis-acting elements in the first intron, and their cognate DNA-binding proteins, mediate transcription of the alpha 1(I) collagen gene in some cells, such as dermal fibroblasts, but not in tendon cells or osteoblasts. Moreover, regions of the gene not included in the sequence, -2300 to +1440, appear to be required for transcription in tissues such as skeletal muscle and perichondrium.

scholarly journals Tissue-specific expression in the salivary glands of transgenic mice

1992 ◽  
Vol 20 (9) ◽  
pp. 2249-2255 ◽  
Author(s):  
Thomas R. Mikkelsen ◽  
Jakob Brandt ◽  
H.Jakob Larsen ◽  
Birte B. Larsen ◽  
Knud Poulsen ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Salivary Glands ◽  
Specific Expression ◽  
Tissue Specific ◽  
Tissue Specific Expression

Tissue-specific regulation of mouse hepatocyte nuclear factor 4 expression

1994 ◽  
Vol 14 (11) ◽  
pp. 7276-7284
Author(s):  
W Zhong ◽  
J Mirkovitch ◽  
J E Darnell
Keyword(s):  
Transcription Factor ◽  
Transgenic Mice ◽  
Transient Transfection ◽  
Nuclear Factor ◽  
Hepatocyte Nuclear Factor ◽  
Distal Enhancer ◽  
Specific Expression ◽  
Tissue Specific ◽  
Hypersensitive Sites ◽  
Hepatocyte Nuclear Factor 4

Hepatocyte nuclear factor 4 (HNF-4) is a liver-enriched transcription factor and a member of the steroid hormone receptor superfamily. HNF-4 is required for the hepatoma-specific expression of HNF-1 alpha, another liver-enriched transcription factor, suggesting the early participation of HNF-4 in development. To prepare for further study of HNF-4 in development, the tissue-specific expression of the mouse HNF-4 gene was studied by analyzing the promoter region for required DNA elements. DNase-hypersensitive sites in the gene in liver and kidney tissues were found in regions both distal and proximal to the RNA start that were absent in tissues in which HNF-4 expression did not occur. By use of reporter constructs in transient-transfection assays and with transgenic mice, a region sufficient to drive liver-specific expression of HNF-4 was identified. While an HNF-1 binding site between bp -98 and -68 played an important role in the hepatoma-specific promoter activity of HNF-4 in transient-transfection assays, it was not sufficient for the liver-specific expression of a reporter gene in transgenic mice. Distal enhancer elements indicated by the presence of DNase I-hypersensitive sites at kb -5.5 and -6.5, while not functional in transient-transfection assays, were required for the correct expression of the mouse HNF-4 gene in animals.

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