Tissue- and development-specific expression in transgenic mice of a type I procollagen (COL1A1) minigene construct with 2.3 kb of the promoter region and 2 kb of the 3'-flanking region. Specificity is independent of the putative regulatory sequences in the first intron

We have identified two blocks of regulatory sequences located in the 5'-flanking region and the first intron of the mouse alpha 1 type I collagen (COL1A1) gene. Both blocks were found to contain positive as well as negative regulatory elements. Sequences located within 222 base pairs upstream of the transcription start site showed a strong stimulatory effect on the COL1A1 promoter and were sufficient for tissue-specific regulation of the COL1A1 gene. The combined upstream and intron regulatory sequences showed a marked inhibition of COL1A1 promoter activity in fibroblasts. This finding suggests that additional, more remote regulatory sequences may be required for establishing the high level of activity of the endogenous COL1A1 gene in fibroblastoid cells.

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Regulatory elements in the 5'-flanking region and the first intron contribute to transcriptional control of the mouse alpha 1 type I collagen gene.

Molecular and Cellular Biology ◽

10.1128/mcb.9.5.2224 ◽

1989 ◽

Vol 9 (5) ◽

pp. 2224-2227 ◽

Cited By ~ 72

Author(s):

R A Rippe ◽

S I Lorenzen ◽

D A Brenner ◽

M Breindl

Keyword(s):

Transcriptional Control ◽

Type I Collagen ◽

Regulatory Elements ◽

Regulatory Sequences ◽

Type I ◽

Col1a1 Gene ◽

First Intron ◽

Flanking Region ◽

Specific Regulation ◽

High Level

We have identified two blocks of regulatory sequences located in the 5'-flanking region and the first intron of the mouse alpha 1 type I collagen (COL1A1) gene. Both blocks were found to contain positive as well as negative regulatory elements. Sequences located within 222 base pairs upstream of the transcription start site showed a strong stimulatory effect on the COL1A1 promoter and were sufficient for tissue-specific regulation of the COL1A1 gene. The combined upstream and intron regulatory sequences showed a marked inhibition of COL1A1 promoter activity in fibroblasts. This finding suggests that additional, more remote regulatory sequences may be required for establishing the high level of activity of the endogenous COL1A1 gene in fibroblastoid cells.

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Differential regulation of the rat phosphoenolpyruvate carboxykinase gene expression in several tissues of transgenic mice.

Molecular and Cellular Biology ◽

10.1128/mcb.12.3.1396 ◽

1992 ◽

Vol 12 (3) ◽

pp. 1396-1403 ◽

Cited By ~ 40

Author(s):

C L Eisenberger ◽

H Nechushtan ◽

H Cohen ◽

M Shani ◽

L Reshef

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Transgenic Mice ◽

Phosphoenolpyruvate Carboxykinase ◽

Developmental Regulation ◽

Regulation Of Gene Expression ◽

Regulatory Sequences ◽

Specific Expression ◽

Endogenous Gene ◽

Flanking Region

The selective expression of a unique copy gene in several mammalian tissues has been approached by studying the regulatory sequences needed to control expression of the rat phosphoenolpyruvate carboxykinase (PEPCK) gene in transgenic mice. A transgene containing the entire PEPCK gene, including 2.2 kb of the 5'-flanking region and 0.5 kb of the 3'-flanking region, exhibits tissue-specific expression in the liver, kidney, and adipose tissue, as well as the hormonal and developmental regulation inherent to endogenous gene expression. Deletions of the 5'-flanking region of the gene have shown the need for sequences downstream of position -540 of the PEPCK gene for expression in the liver and sequences downstream of position -362 for expression in the kidney. Additional sequences upstream of position -540 (up to -2200) are required for expression in adipose tissue. In addition, the region containing the glucocorticoid-responsive elements of the gene used by the kidney was identified. This same sequence was found to be needed specifically for developmental regulation of gene expression in the kidney and, together with upstream sequences, in the intestine. The apparently distinct sequence requirements in the various tissues indicate that the tissues use different mechanisms for expression of the same gene.

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Tissue-specific Expression of the Gene for Type I Procollagen (COL1A1) in Transgenic Mice

Journal of Biological Chemistry ◽

10.1074/jbc.270.16.9622 ◽

1995 ◽

Vol 270 (16) ◽

pp. 9622-9629 ◽

Cited By ~ 23

Author(s):

Boris P. Sokolov ◽

Leena Ala-Kokko ◽

Rohini Dhulipala ◽

Machiko Arita ◽

Jaspal S. Khillan ◽

...

Keyword(s):

Transgenic Mice ◽

Type I ◽

Specific Expression ◽

Tissue Specific ◽

Tissue Specific Expression ◽

Type I Procollagen

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Differential regulation of the rat phosphoenolpyruvate carboxykinase gene expression in several tissues of transgenic mice

Molecular and Cellular Biology ◽

10.1128/mcb.12.3.1396-1403.1992 ◽

1992 ◽

Vol 12 (3) ◽

pp. 1396-1403

Author(s):

C L Eisenberger ◽

H Nechushtan ◽

H Cohen ◽

M Shani ◽

L Reshef

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Transgenic Mice ◽

Phosphoenolpyruvate Carboxykinase ◽

Developmental Regulation ◽

Regulation Of Gene Expression ◽

Regulatory Sequences ◽

Specific Expression ◽

Endogenous Gene ◽

Flanking Region

The selective expression of a unique copy gene in several mammalian tissues has been approached by studying the regulatory sequences needed to control expression of the rat phosphoenolpyruvate carboxykinase (PEPCK) gene in transgenic mice. A transgene containing the entire PEPCK gene, including 2.2 kb of the 5'-flanking region and 0.5 kb of the 3'-flanking region, exhibits tissue-specific expression in the liver, kidney, and adipose tissue, as well as the hormonal and developmental regulation inherent to endogenous gene expression. Deletions of the 5'-flanking region of the gene have shown the need for sequences downstream of position -540 of the PEPCK gene for expression in the liver and sequences downstream of position -362 for expression in the kidney. Additional sequences upstream of position -540 (up to -2200) are required for expression in adipose tissue. In addition, the region containing the glucocorticoid-responsive elements of the gene used by the kidney was identified. This same sequence was found to be needed specifically for developmental regulation of gene expression in the kidney and, together with upstream sequences, in the intestine. The apparently distinct sequence requirements in the various tissues indicate that the tissues use different mechanisms for expression of the same gene.

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Tissue-specific expression of human CD4 in transgenic mice

Molecular and Cellular Biology ◽

10.1128/mcb.13.5.2952-2958.1993 ◽

1993 ◽

Vol 13 (5) ◽

pp. 2952-2958

Author(s):

F P Gillespie ◽

L Doros ◽

J Vitale ◽

C Blackwell ◽

J Gosselin ◽

...

Keyword(s):

T Cell ◽

Transgenic Mice ◽

Cell Line ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Specific Expression ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

Flanking Region

The gene for the human CD4 glycoprotein, which serves as the receptor for human immunodeficiency virus type 1, along with approximately 23 kb of sequence upstream of the translational start site, was cloned. The ability of 5' flanking sequences to direct tissue-specific expression was tested in cell culture and in transgenic mice. A 5' flanking region of 6 kb was able to direct transcription of the CD4 gene in NIH 3T3 cells but did not result in detectable expression in the murine T-cell line EL4 or in four lines of transgenic mice. A larger 5' flanking region of approximately 23 kb directed high-level CD4 transcription in the murine T-cell line EL4 and in three independent lines of transgenic mice. Human CD4 expression in all tissues analyzed was tightly correlated with murine CD4 expression; the highest levels of human CD4 RNA expression were found in the thymus and spleen, with relatively low levels detected in other tissues. Expression of human CD4 protein in peripheral blood mononuclear cells was examined by flow cytometry in these transgenic animals and found to be restricted to the murine CD4+ subset of lymphocytes. Human CD4 protein, detected with an anti-human CD4 monoclonal antibody, was present on the surface of 45 to 50% of the peripheral blood mononuclear cells from all transgenic lines.

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Beta-MHC and SMLC1 transgene induction in overloaded skeletal muscle of transgenic mice

AJP Cell Physiology ◽

10.1152/ajpcell.1996.270.4.c1111 ◽

1996 ◽

Vol 270 (4) ◽

pp. C1111-C1121 ◽

Cited By ~ 15

Author(s):

J. L. Wiedenman ◽

I. Rivera-Rivera ◽

D. Vyas ◽

G. Tsika ◽

L. Gao ◽

...

Keyword(s):

Transgenic Mice ◽

Troponin C ◽

Type I ◽

Initial Attempt ◽

Specific Expression ◽

Slow Type ◽

Transgenic Lines ◽

Fast Twitch Muscle ◽

Mechanical Overload

The hypertrophic responses of white fast-twitch muscle to mechanical overload has been investigated using transgenic mice. After 7 wk of overload, endogenous beta-myosin heavy chain (MHC) and slow myosin light chain 1 and 2 (SMLC1, SMLC2) protein were increased in the overloaded plantaris (OP) muscle compared with sham-operated control plantaris (CP)muscle. Concurrently, the levels of endogenous beta-MHC, SMLC1, SMLC2, and cardiac/slow troponin C (CTnC) mRNA transcripts were significantly increased in OP muscles, whereas skeletal troponin C (sTnC) mRNA transcript levels decreased. As an initial attempt to locate DNA sequence(s) that governs beta-MHC induction in response to mechanical overload, multiple independent transgenic lines harboring four different human beta-MHC transgenes (beta 1286, beta 988, beta 450, beta 141) were generated. Except for transgene beta 141, muscle-specific expression and induction (3- to 22-fold) in OP muscles were observed by measuring chloramphenicol acetyltransferase activity (CAT assay). Induction of a SMLC1 transgene (3920SMLC1) in OP muscles was also observed. Collectively, these in vivo data provide evidence that 1) a mechanical overload inducible element(s) is located between nucleotides -450 and +120 of the human beta-MHC transgene, 2) 3,900 bp of 5' sequence is sufficient to confer mechanical overload induction of a SMLC1 transgene, and 3) the increased expression of slow/type I isomyosin (beta-MHC, SMLC1, SMLC2) in response to mechanical overload is regulated, in part, transcriptionally.

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beta-MHC transgene expression in suspended and mechanically overloaded/suspended soleus muscle of transgenic mice

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1997.272.5.r1552 ◽

1997 ◽

Vol 272 (5) ◽

pp. R1552-R1561 ◽

Cited By ~ 22

Author(s):

J. J. McCarthy ◽

A. M. Fox ◽

G. L. Tsika ◽

L. Gao ◽

R. W. Tsika

Keyword(s):

Transgenic Mice ◽

Soleus Muscle ◽

Transgene Expression ◽

Specific Activity ◽

Fiber Type ◽

Weight Bearing ◽

Troponin C ◽

Hindlimb Suspension ◽

Regulatory Sequences ◽

Type I

Non-weight-bearing (NWB) activity [space flight and hindlimb suspension (HS)] results in the loss of soleus muscle mass, a slow-to-fast fiber-type conversion, and decreased beta-myosin heavy chain (beta-MHC) protein and mRNA expression. To identify beta-MHC promoter sequences required for decreased beta-MHC expression in response to HS, we have modified an existing noninvasive hindlimb unweighting model to accommodate the use of (transgenic) mice. After 2 wk of HS, body and muscle (soleus > gastrocnemius > plantaris) weights were decreased as was the proportion of histochemically classified type I fibers in HS soleus muscle. Northern blot analysis revealed decreases in endogenous mRNA representing beta-MHC, slow myosin light chain 1 and 2, and cardiac/slow troponin C, whereas those representing skeletal troponin C, muscle creatine kinase, and glyceraldehyde-3-phosphate dehydrogenase increased. Protein extracts prepared from HS soleus (SS) muscle of mice harboring transgenes comprised of 5.6 or 0.6 kilobase of wild type (wt) mouse beta-MHC promoter (beta 5.6 wt, beta 0.6wt) and those carrying the simultaneous mutation (mut) of the MCAT, C-rich, and beta e3 subregions (beta 5.6mut3, beta 0.6mut3) revealed decreases in chloramphenicol acetyltransferase (CAT) specific activity relative to respective controls. Decreased CAT mRNA was observed for transgene beta 5.6mut3, line 85. Two weeks of the simultaneous imposition of mechanical overload (synergist ablation) and HS (MOV/HS) countermanded the loss in absolute and normalized SS weight but did not decrease beta 0.6wt transgene expression. These transgenic results demonstrate that regulatory sequences within a 600-base pair beta-MHC promoter are sufficient to direct decreased transcription of beta-MHC transgenes after 2 wk of HS.

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