Universität zu Köln

Kuri, Paola ORCID: 0000-0003-4219-8417, Ellwanger, Kornelia, Kufer, Thomas A., Leptin, Maria ORCID: 0000-0001-7097-348X and Bajoghli, Baubak ORCID: 0000-0002-7368-7523 (2017). A high-sensitivity bi-directional reporter to monitor NF-kappa B activity in cell culture and zebrafish in real time. J. Cell Sci., 130 (3). S. 648 - 658. CAMBRIDGE: COMPANY OF BIOLOGISTS LTD. ISSN 1477-9137

Full text not available from this repository.

Abstract

Nuclear factor (NF)-kappa B transcription factors play major roles in numerous biological processes including development and immunity. Here, we engineered a novel bi-directional NF-kappa B-responsive reporter, pSGNluc, in which a high-affinity NF-kappa B promoter fragment simultaneously drives expression of luciferase and GFP. Treatment with TNF alpha (also known as TNF) induced a strong, dosedependent luciferase signal in cell culture. The degree of induction over background was comparable to that of other NF-kappa B-driven luciferase reporters, but the absolute level of expression was at least 20-fold higher. This extends the sensitivity range of otherwise difficult assays mediated exclusively by endogenously expressed receptors, as we show for Nod1 signaling in HEK293 cells. To measure NF-kappa B activity in the living organism, we established a transgenic zebrafish line carrying the pSGNluc construct. Live in toto imaging of transgenic embryos revealed the activation patterns of NF-kappa B signaling during embryonic development and as responses to inflammatory stimuli. Taken together, by integrating qualitative and quantitative NF-kappa B reporter activity, pSGNluc is a valuable tool for studying NF-kappa B signaling at high spatiotemporal resolution in cultured cells and living animals that goes beyond the possibilities provided by currently available reporters.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Kuri, Paola UNSPECIFIED orcid.org/0000-0003-4219-8417 UNSPECIFIED Ellwanger, Kornelia UNSPECIFIED UNSPECIFIED UNSPECIFIED Kufer, Thomas A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Leptin, Maria UNSPECIFIED orcid.org/0000-0001-7097-348X UNSPECIFIED Bajoghli, Baubak UNSPECIFIED orcid.org/0000-0002-7368-7523 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-240851
DOI:	10.1242/jcs.196485
Journal or Publication Title:	J. Cell Sci.
Volume:	130
Number:	3
Page Range:	S. 648 - 658
Date:	2017
Publisher:	COMPANY OF BIOLOGISTS LTD
Place of Publication:	CAMBRIDGE
ISSN:	1477-9137
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Institute for Genetics
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language IN-VIVO; HIGH-THROUGHPUT; BACTERIAL PEPTIDOGLYCAN; GENE-EXPRESSION; IMMUNE-SYSTEM; ACTIVATION; NOD1; ALPHA; MICE; INFLAMMATION Multiple languages Cell Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/24085