Nature Methods
2, 607 - 614 (2005)
Published online: 21 July 2005; | doi:10.1038/nmeth779
Real-time imaging of ligand-induced IKK activation in intact cells and in living miceShimon Gross1
& David Piwnica-Worms1, 21
Molecular Imaging Center, Mallinckrodt Institute of Radiology, Washington University School of Medicine, 510 S. Kingshighway Blvd., Box 8225, St. Louis, Missouri 63110, USA. 2
Department of Molecular Biology and Pharmacology, Washington University School of Medicine, 510 S. Kingshighway Blvd., Box 8225, St. Louis, Missouri 63110, USA.
Correspondence should be addressed to David Piwnica-Worms piwnica-wormsd@mir.wustl.edu The transcription factor NF- B is a key regulator of cellular activation, proliferation and apoptosis. Defects in the NF-B pathway contribute to a broad array of malignant, neurodegenerative and chronic inflammatory diseases. IKK-dependent IB degradation by the 26S proteasome is a critical NF-B regulatory control point, which is emerging as an important target for drug development. To directly monitor regulation of IKK activation in intact organisms, we engineered an IB−firefly luciferase (IB-FLuc) fusion reporter. In cultured cells and living animals, the reporter provided a continuous, noninvasive readout of the kinetics of ligand-induced IKK activation and the pharmacodynamics of selective inhibitors of both IKK and the 26S proteasome. This IB-FLuc reporter now permits continuous readout of IKK activation in vivo, facilitates development and validation of target-specific therapeutics, and complements conventional NF-B transcriptional reporters for more complete temporal and regional investigations of the NF-B signaling pathway in health and disease.
MORE ARTICLES LIKE THIS These links to content published by NPG are automatically generated.
|
B
-EGFP plasmid (BD Bioscience)
FLuc (Stratagene)
