An immune response after intraocular administration of an adenoviral vector containing a beta galactosidase reporter gene slows retinal degeneration in the rd mouse

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.

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Protein phosphatase 2A potentiates activity of promoters containing AP-1-binding elements

Molecular and Cellular Biology ◽

10.1128/mcb.13.4.2104-2112.1993 ◽

1993 ◽

Vol 13 (4) ◽

pp. 2104-2112

Author(s):

A S Alberts ◽

T Deng ◽

A Lin ◽

J L Meinkoth ◽

A Schönthal ◽

...

Keyword(s):

Gene Expression ◽

Reporter Gene ◽

Protein Phosphatase ◽

Signal Transduction Pathways ◽

Marker Genes ◽

Direct Mechanism ◽

Phosphatase 2A ◽

Beta Galactosidase ◽

Regulatory Sites

The involvement of serine/threonine protein phosphatases in signaling pathways which modulate the activity of the transcription factor AP-1 was examined. Purified protein phosphatase types 1 (PP1) and 2A (PP2A) were microinjected into cell lines containing stably transfected lacZ marker genes under the control of an enhancer recognized by AP-1. Microinjection of PP2A potentiated serum-stimulated beta-galactosidase expression from the AP-1-regulated promoter. Similarly, transient expression of the PP2A catalytic subunit with c-Jun resulted in a synergistic transactivation of an AP-1-regulated reporter gene. PP2A, but not PP1, potentiated serum-induced c-Jun expression, which has been previously shown to be autoregulated by AP-1 itself. Consistent with these results, PP2A dephosphorylated c-Jun on negative regulatory sites in vitro, suggesting one possible direct mechanism for the effects of PP2A on AP-1 activity. Microinjection of PP2A had no effect on cyclic AMP (cAMP)-induced expression of a reporter gene containing a cAMP-regulated promoter, while PP1 injection abolished cAMP-induced gene expression. Taken together, these results suggest a specific role for PP2A in signal transduction pathways that regulate AP-1 activity and c-Jun expression.

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Biodistribution of an adenoviral vector carrying the luciferase reporter gene following intravesical or intravenous administration to a mouse

Cancer Gene Therapy ◽

10.1038/sj.cgt.7700090 ◽

1999 ◽

Vol 6 (4) ◽

pp. 367-372 ◽

Cited By ~ 90

Author(s):

Mark Wood ◽

Paul Perrotte ◽

Eric Onishi ◽

Mary E Harper ◽

Colin Dinney ◽

...

Keyword(s):

Reporter Gene ◽

Intravenous Administration ◽

Adenoviral Vector ◽

Luciferase Reporter ◽

Luciferase Reporter Gene

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Protein phosphatase 2A potentiates activity of promoters containing AP-1-binding elements.

Molecular and Cellular Biology ◽

10.1128/mcb.13.4.2104 ◽

1993 ◽

Vol 13 (4) ◽

pp. 2104-2112 ◽

Cited By ~ 50

Author(s):

A S Alberts ◽

T Deng ◽

A Lin ◽

J L Meinkoth ◽

A Schönthal ◽

...

Keyword(s):

Gene Expression ◽

Reporter Gene ◽

Protein Phosphatase ◽

Signal Transduction Pathways ◽

Marker Genes ◽

Direct Mechanism ◽

Phosphatase 2A ◽

Beta Galactosidase ◽

Regulatory Sites

The involvement of serine/threonine protein phosphatases in signaling pathways which modulate the activity of the transcription factor AP-1 was examined. Purified protein phosphatase types 1 (PP1) and 2A (PP2A) were microinjected into cell lines containing stably transfected lacZ marker genes under the control of an enhancer recognized by AP-1. Microinjection of PP2A potentiated serum-stimulated beta-galactosidase expression from the AP-1-regulated promoter. Similarly, transient expression of the PP2A catalytic subunit with c-Jun resulted in a synergistic transactivation of an AP-1-regulated reporter gene. PP2A, but not PP1, potentiated serum-induced c-Jun expression, which has been previously shown to be autoregulated by AP-1 itself. Consistent with these results, PP2A dephosphorylated c-Jun on negative regulatory sites in vitro, suggesting one possible direct mechanism for the effects of PP2A on AP-1 activity. Microinjection of PP2A had no effect on cyclic AMP (cAMP)-induced expression of a reporter gene containing a cAMP-regulated promoter, while PP1 injection abolished cAMP-induced gene expression. Taken together, these results suggest a specific role for PP2A in signal transduction pathways that regulate AP-1 activity and c-Jun expression.

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Choice lines of Drosophila melanogaster with reporter system sensitive to low doses of mutagens

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v21.811 ◽

1970 ◽

Vol 21 ◽

pp. 77-81

Author(s):

Ya. S. Yasinskyi ◽

L. V. Omelianchuk ◽

O. V. Zhuk ◽

I. A. Kozeretska

Keyword(s):

Drosophila Melanogaster ◽

Ionizing Radiation ◽

Reporter Gene ◽

Low Dose ◽

Gene Activation ◽

Fruit Fly ◽

Lacz Reporter ◽

Reporter System ◽

Small Doses ◽

Beta Galactosidase

Aim. There are many methods of assessment of mutagens, but the effects of low-dose mutagens have been overlooked. In this study we try to select lines of Drosophila melanogaster with high-sensitive to mutagen reporter gene. We used D. melanogaster lines with enhancer trap P {lacW} which produces beta-galactosidase gene in response to activation induced reparation. Methods. Ionizing radiation in dose of 100 roentgens were used as a mutagen. We extracted salivary glands of the third instar of the fruit fly larvae. The sample was fixed with 0.75 % glutaraldehyde and stained by 12-hour incubation with 0.2 % X-Gal at 37°C. Activity of the reporter gene was determined by microscope. Results. Two of the three lines are not shown according to the activation of the reporter gene from the resulting dose of ionizing radiation. In line y [1] w [67c23]; P {w [+ mC] = lacW} mus209 [k00704] / CyO (#10361) activation of the reporter gene occurred in some nuclei salivary gland after radiation. Conclusions. The reporter gene P{lacW} may be used for the detection of small doses of mutagens. Larvae of line #10361 has reporter gene activation in response to ionizing radiation. Keywords: mutagenic factors, Drosophila melanogaster, LacZ reporter, radiation.

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Modulation of the Immune Response to the Severe Acute Respiratory Syndrome Spike Glycoprotein by Gene-Based and Inactivated Virus Immunization

Journal of Virology ◽

10.1128/jvi.79.22.13915-13923.2005 ◽

2005 ◽

Vol 79 (22) ◽

pp. 13915-13923 ◽

Cited By ~ 27

Author(s):

Wing-pui Kong ◽

Ling Xu ◽

Konrad Stadler ◽

Jeffrey B. Ulmer ◽

Sergio Abrignani ◽

...

Keyword(s):

Immune Response ◽

Severe Acute Respiratory Syndrome ◽

Immune Responses ◽

Adenoviral Vector ◽

T Cell Response ◽

Immune Recognition ◽

Humoral Immune ◽

Humoral Immune Responses ◽

Viral Particles ◽

Stimulation Of

ABSTRACT Although the initial isolates of the severe acute respiratory syndrome (SARS) coronavirus (CoV) are sensitive to neutralization by antibodies through their spike (S) glycoprotein, variants of S have since been identified that are resistant to such inhibition. Optimal vaccine strategies would therefore make use of additional determinants of immune recognition, either through cellular or expanded, cross-reactive humoral immunity. Here, the cellular and humoral immune responses elicited by different combinations of gene-based and inactivated viral particles with various adjuvants have been assessed. The T-cell response was altered by different prime-boost immunizations, with the optimal CD8 immunity induced by DNA priming and replication-defective adenoviral vector boosting. The humoral immune response was enhanced most effectively through the use of inactivated virus with adjuvants, either MF59 or alum, and was associated with stimulation of the CD4 but not the CD8 response. The use of inactivated SARS virus with MF59 enhanced the CD4 and antibody response even after gene-based vaccination. Because both cellular and humoral immune responses are generated by gene-based vaccination and inactivated viral boosting, this strategy may prove useful in the generation of SARS-CoV vaccines.

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