Conditional and inducible transgene expression in endothelial and hematopoietic cells using Cre/loxP and tetracycline-off systems

Liu, Ju; Deutsch, Urban; Fung, Iris; Lobe, Corrinne G (2014). Conditional and inducible transgene expression in endothelial and hematopoietic cells using Cre/loxP and tetracycline-off systems. Experimental and therapeutic medicine, 8(5), pp. 1351-1356. Spandidos Publ. 10.3892/etm.2014.1965

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In the present study, the tetracycline-off and Cre/loxP systems were combined to gain temporal and spatial control of transgene expression. Mice were generated that carried three transgenes: Tie2-tTA, tet-O-Cre and either the ZEG or ZAP reporter. Tie2-tTA directs expression of tetracycline-controlled transactivator (tTA) in endothelial and hematopoietic cells under the control of the Tie2 promoter. Tet-O-Cre produces Cre recombinase from a minimal promoter containing the tet-operator (tetO). ZEG or ZAP contains a strong promoter and a loxP-flanked stop sequence, followed by an enhanced green fluorescence protein (EGFP) or human placental alkaline phosphatase (hPLAP) reporter. In the presence of tetracycline, the tTA transactivator produced by Tie-2-tTA is disabled and Cre is not expressed. In the absence of tetracycline, the tTA binds tet-O-Cre to drive the expression of Cre, which recombines the loxP sites of the ZEG or ZAP transgene and results in reporter gene expression. In the present study, the expression of the ZEG or ZAP reporter genes in embryos and adult animals with and without tetracycline treatment was examined. In the presence of tetracycline, no reporter gene expression was observed. When tetracycline was withdrawn, Cre excision was activated and the reporter genes were detected in endothelial and hematopoietic cells. These results demonstrate that this system may be used to bypass embryonic lethality and access adult phenotypes.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Theodor Kocher Institute

UniBE Contributor:

Deutsch, Urban

Subjects:

600 Technology > 610 Medicine & health

ISSN:

1792-0981

Publisher:

Spandidos Publ.

Language:

English

Submitter:

Ursula Zingg-Zünd

Date Deposited:

07 Apr 2015 13:18

Last Modified:

07 Apr 2015 13:19

Publisher DOI:

10.3892/etm.2014.1965

PubMed ID:

25289022

Uncontrolled Keywords:

Cre/loxP system, endothelial cells, hematopoietic cell, tetracycline, transgenic mice

BORIS DOI:

10.7892/boris.66367

URI:

https://boris.unibe.ch/id/eprint/66367

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