Kuri, Paola ORCID: 0000-0003-4219-8417, Ellwanger, Kornelia, Kufer, Thomas A., Leptin, Maria 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

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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:
CreatorsEmailORCIDORCID Put Code
Kuri, PaolaUNSPECIFIEDorcid.org/0000-0003-4219-8417UNSPECIFIED
Ellwanger, KorneliaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Kufer, Thomas A.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Leptin, MariaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Bajoghli, BaubakUNSPECIFIEDorcid.org/0000-0002-7368-7523UNSPECIFIED
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: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
IN-VIVO; HIGH-THROUGHPUT; BACTERIAL PEPTIDOGLYCAN; GENE-EXPRESSION; IMMUNE-SYSTEM; ACTIVATION; NOD1; ALPHA; MICE; INFLAMMATIONMultiple languages
Cell BiologyMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/24085

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